Platelet ADP receptor inhibitors

ABSTRACT

Novel compounds of formulae (I) to (VIII), which more particularly include sulfonylurea derivatives, sulfonylthiourea derivatives, sulfonylguanidine derivatives, sulfonylcyanoguanidine derivatives, thioacylsulfonamide derivatives, and acylsulfonamide derivatives which are effective platelet ADP receptor inhibitors. These derivatives may be used in various pharmaceutical compositions, and are particularly effective for the prevention and/or treatment of cardiovascular diseases, particularly those diseases related to thrombosis. The invention also relates to a method for preventing or treating thrombosis in a mammal comprising the step of administering a therapeutically effective amount of a compound of formulae (I) to (VIII), or a pharmaceutically acceptable salt thereof.

The present application is a continuation of application Ser. No.09/920,325, filed Aug. 2, 2001, which is a continuation-in-part (CIP) ofU.S. application Ser. No. 09/775,812, filed Feb. 5, 2001, now abandoned;claiming benefit of 60/180,208, filed Feb. 4, 2000; 60/202,072, filedMay 5, 2000; 60/230,447, filed Sep. 6, 2000 and a CIP of internationalapplication PCT/US01/03585, filed Feb. 5, 2001, designating the U.S.,all of which are incorporated by reference.

FIELD OF THE INVENTION

The invention relates to novel compounds of formula (I), formula (II),formula (III), formula (IV), formula (V), formula (VI), formula (VII)and formula (VIII) (hereinafter referred to as “formulae (I)–(VIII)”),which more particularly include sulfonylurea derivatives,sulfonylthiourea derivatives, sulfonylguanidine derivatives,sulfonylcyanoguanidine derivatives, thioacylsulfonamide derivatives, andacylsulfonamide derivatives which are effective platelet ADP receptorinhibitors. These derivatives may be used in various pharmaceuticalcompositions, and are particularly effective for the prevention and/ortreatment of cardiovascular diseases, particularly those diseasesrelated to thrombosis.

DESCRIPTION OF THE RELATED ART

Thrombotic complications are a major cause of death in theindustrialized world. Examples of these complications include acutemyocardial infarction, unstable angina, chronic stable angina, transientischemic attacks, strokes, peripheral vascular disease,preeclampsia/eclampsia, deep venous thrombosis, embolism, disseminatedintravascular coagulation and thrombotic cytopenic purpura. Thromboticand restenotic complications also occur following invasive procedures,e.g., angioplasty, carotid endarterectomy, post CABG (coronary arterybypass graft) surgery, vascular graft surgery, stent placements andinsertion of endovascular devices and protheses. It is generally thoughtthat platelet aggregates play a critical role in these events. Bloodplatelets, which normally circulate freely in the vasculature, becomeactivated and aggregate to form a thrombus with disturbed blood flowcaused by ruptured atherosclerotic lesions or by invasive treatmentssuch as angioplasty, resulting in vascular occlusion. Plateletactivation can be initiated by a variety of agents, e.g., exposedsubendothelial matrix molecules such as collagen, or by thrombin whichis formed in the coagulation cascade.

An important mediator of platelet activation and aggregation is ADP(adenosine 5′-diphosphate) which is released from blood platelets in thevasculature upon activation by various agents, such as collagen andthrombin, and from damaged blood cells, endothelium or tissues.Activation by ADP results in the recruitment of more platelets andstabilization of existing platelet aggregates. Platelet ADP receptorsmediating aggregation are activated by ADP and some of its derivativesand antagonized by ATP (adenosine 5′-triphosphate) and some of itsderivatives (Mills, D. C. B. (1996) Thromb. Hemost. 76:835–856).Therefore, platelet ADP receptors are members of the family of P2receptors activated by purine and/or pyrimidine nucleotides (King, B.F., Townsend-Nicholson, A. & Burnstock, G. (1998) Trends Pharmacol. Sci.19:506–514).

Recent pharmacological data using selective antagonists suggests thatADP-dependent platelet aggregation requires activation of at least twoADP receptors (Kunapuli, S. P. (1998), Trends Pharmacol. Sci.19:391–394; Kunapuli, S. P. & Daniel, J. L. (1998) Biochem. J.336:513–523; Jantzen, H. M. et al. (1999) Thromb. Hemost. 81: 111–117).One receptor appears to be identical to the cloned P2Y₁ receptor,mediates phospholipase C activation and intracellular calciummobilization and is required for platelet shape change. The secondplatelet ADP receptor important for aggregation mediates inhibition ofadenylyl cyclase. Molecular cloning of the gene or cDNA for thisreceptor (P2Y₁₂) has recently been reported (Hollopeter, G. et. al.(2001) Nature 409:202–207). Based on its pharmacological and signalingproperties this receptor has been previously termed P2Y_(ADP) (Fredholm,B. B. et al. (1997) TIPS 18:79–82), P2T_(AC) (Kunapuli, S. P. (1998),Trends Pharmacol. Sci. 19:391–394) or P2Ycyc (Hechler, B. et al. (1998)Blood 92, 152–159).

Various directly or indirectly acting synthetic inhibitors ofADP-dependent platelet aggregation with antithrombotic activity havebeen reported. The orally active antithrombotic thienopyridinesticlopidine and clopidogrel inhibit ADP-induced platelet aggregation,binding of radiolabeled ADP receptor agonist 2-methylthioadenosine5′-diphosphate to platelets, and other ADP-dependent events indirectly,probably via formation of an unstable and irreversible acting metabolite(Quinn, M. J. & Fitzgerald, D. J. (1999) Circulation 100:1667–1667).Some purine derivatives of the endogenous antagonist ATP, e.g., AR-C(formerly FPL or ARL) 67085MX and AR-C69931MX, are selective plateletADP receptor antagonists which inhibit ADP-dependent plateletaggregation and are effective in animal thrombosis models (Humphries etal. (1995), Trends Pharmacol. Sci. 16, 179; Ingall, A. H. et al. (1999)J. Med. Chem. 42, 213–230). Novel triazolo [4,5-d] pyrimidine compoundshave been disclosed as P_(2T)—antagonists (WO 99/05144). Tricycliccompounds as platelet ADP receptor inhibitors have also been disclosedin WO 99/36425. The target of these antithrombotic compounds appears tobe the platelet ADP receptor mediating inhibition of adenylyl cyclase.

Despite these compounds, there exists a need for more effective plateletADP receptor inhibitors. In particular, there is a need for platelet ADPreceptor inhibitors having antithrombotic activity that are useful inthe prevention and/or treatment of cardiovascular diseases, particularlythose related to thrombosis.

SUMMARY OF THE INVENTION

The invention provides compounds of formula (I), formula (II), formula(III), formula (IV), formula (V), formula (VI), formula (VII) andformula (VIII):

-   A is selected from the group consisting of aryl, substituted aryl,    heteroaryl, substituted heteroaryl, alkylaryl, and alkylheteroaryl.-   W is selected from the group consisting of aryl, substituted aryl,    heteroaryl, and substituted heteroaryl.-   E is selected from the group consisting of H, —C₁–C₈ alkyl,    polyhaloalkyl, —C₃₋₈cycloalkyl, aryl, alkylaryl, substituted aryl,    heteroaryl, and substituted heteroaryl.-   D is selected from the group consisting of NR¹-(C═O)—R², —O—R¹;

wherein:

-   R¹ is independently selected from the group consisting of:

H, C₁–C₈ alkyl, polyhaloalkyl, —C₃₋₈-cycloalkyl, aryl, alkylaryl,substituted aryl, heteroaryl, substituted heteroaryl, —(C═O)—C₁–C₈alkyl, —(C═O)-aryl, —(C═O)-substituted aryl, —(C═O)-heteroaryl and—(C═O)-substituted heteroaryl;

-   R² is independently selected from the group consisting of aryl,    substituted aryl, heteroaryl, substituted heteroaryl, or-   R¹ and R² can be direct linked or can be indirectly linked through a    carbon chain that is from 1 to about 8 carbon atoms in length,-   n is 0–4,-   m is 0 or 1,-   y is 0–4 and-   Q is independently C or N, with the proviso that when Q is a ring    carbon atom, each ring carbon atom is independenty substituted by X.-   X is in each case a member independently selected from the group    consisting of:    -   H, halogen, polyhaloalkyl, —OR³, —SR³, —CN, —NO₂, —SO₂R³,        —C₁₋₁₀-alkyl, —C₃₋₈-cycloalkyl, aryl, aryl-substituted by 1–4 R³        groups, amino, amino-C₁₋₈-alkyl, C₁₋₃-acylamino,        C₁₋₃-acylamino-C₁₋₈-alkyl, C₁₋₆-alkylamino, C₁₋₆-alkylamino C₁₋₈        alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl, C₁₋₆        alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl, carboxy-C₁₋₆-alkyl,        C₁₋₃-alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl, carboxy        C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl, and a 5 to 10        membered fused or non-fused aromatic or nonaromatic heterocyclic        ring system, having 1 to 4 heteroatoms independently selected        from N, O, and S, with the proviso that the carbon and nitrogen        atoms, when present in the heterocyclic ring system, are        unsubstituted, mono- or disubstituted independently with 0–2 R⁴        groups.-   R³ and R⁴ are each independently selected from the group consisting    of:    -   H, halogen, —CN, —NO₂, —C₁₋₁₀ alkyl, C₃₋₈-cycloalkyl, aryl,        amino, amino-C₁₋₈-alkyl, C₁₋₃-acylamino,        C₁₋₃-acylamino-C₁₋₈-alkyl, C₁₋₆alkylamino, C₁₋₆-alkylamino C₁₋₈        alkyl, C-6 dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl, C₁₋₆        alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl, carboxy-C₁₋₆-alkyl,        C₁₋₃-alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl,        carboxy-C₁₋₆-alkyloxy, hydroxy, hydroxy-C₁₋₆-alkyl, -thio and        thio-C₁₋₆-alkyl.-   Y is selected from the group consisting of O, S, N—OR⁵, and NR⁵,    wherein R⁵ is selected from the group consisting of:    -   H, C₁₋₁₀ alkyl, C₃₋₈-cycloalkyl, NR², and CN; and-   Z is selected from the group consisting of NR1 and 0.

The invention also covers all pharmaceutically acceptable salts andprodrugs of the compounds of formulae (I)–(VIII).

In another aspect, the invention provides pharmaceutical compositionsfor preventing or treating thrombosis in a mammal containing atherapeutically effective amount of a compound of formulae (I)–(VIII) ora pharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier. The invention further provides a method forpreventing or treating thrombosis in a mammal by administering atherapeutically effective amount of a compound of formulae (I)–(VIII) ora pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

In accordance with the present invention and as used herein, thefollowing terms are defined with the following meanings, unlessexplicitly stated otherwise.

The term “alkenyl” refers to a trivalent straight chain or branchedchain unsaturated aliphatic radical. The term “alkinyl” (or “alkynyl”)refers to a straight or branched chain aliphatic radical that includesat least two carbons joined by a triple bond. If no number of carbons isspecified, alkenyl and alkinyl each refer to radicals having from 2–12carbon atoms.

The term “alkyl” refers to saturated aliphatic groups includingstraight-chain, branched-chain and cyclic groups having the number ofcarbon atoms specified, or if no number is specified, having up to about12 carbon atoms. The term “cycloalkyl” as used herein refers to a mono-,bi-, or tricyclic aliphatic ring having 3 to about 14 carbon atoms andpreferably 3 to about 7 carbon atoms.

The term “C₁–C₆ alkoxy” as used herein refers to an ether moiety wherebythe oxygen is connected to a straight or branched chain of carbon atomsof the number indicated.

The term “mono-C₁–C₆ alkylamino” as used herein refers to an aminomoiety whereby the nitrogen is substituted with one H and one C₁–C₆alkyl substituent, the latter being defined as above.

The term “di-C₁–C₆ alkylamino” as used herein refers to an amino moietywhereby the nitrogen is substituted with two C₁–C₆ alkyl substituents asdefined above.

The term “monoarylamino” as used herein refers to an amino moietywhereby the nitrogen is substituted with one H and one aryl substituent,such as a phenyl, the latter being defined as above.

The term “diarylamino” as used herein refers to an amino moiety wherebythe nitrogen is substituted with two aryl substituents, such as phenyl,the latter being defined as above.

The term “C₁–C₆ alkylsulfonyl” as used herein refers to a dioxosulfurmoiety with the sulfur atom also connected to one C₁–C₆ alkylsubstituent, the latter being defined as above.

The term “C₁–C₆ alkoxycarbonyl” as used herein refers to ahydroxycarbonyl moiety whereby the hydrogen is replaced by a C₁–C₆ alkylsubstituent, the latter being defined as above.

As used herein, the terms “carbocyclic ring structure” and “C₃₋₁₆carbocyclic mono, bicyclic or tricyclic ring structure” or the like areeach intended to mean stable ring structures having only carbon atoms asring atoms wherein the ring structure is a substituted or unsubstitutedmember selected from the group consisting of: a stable monocyclic ringwhich is an aromatic ring (“aryl”) having six ring atoms (“phenyl”); astable monocyclic non-aromatic ring having from 3 to about 7 ring atomsin the ring; a stable bicyclic ring structure having a total of from 7to about 12 ring atoms in the two rings wherein the bicyclic ringstructure is selected from the group consisting of ring structures inwhich both of the rings are aromatic, ring structures in which one ofthe rings is aromatic and ring structures in which both of the rings arenon-aromatic; and a stable tricyclic ring structure having a total offrom about 10 to about 16 atoms in the three rings wherein the tricyclicring structure is selected from the group consisting of: ring structuresin which three of the rings are aromatic, ring structures in which twoof the rings are aromatic and ring structures in which three of therings are non-aromatic. In each case, the non-aromatic rings whenpresent in the monocyclic, bicyclic or tricyclic ring structure mayindependently be saturated, partially saturated or fully saturated.Examples of such carbocyclic ring structures include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl,cyclooctyl, [3.3.0]bicyclooctane, [4.3.0]bicyclononane,[4.4.0]bicyclodecane (decalin), 2.2.2]bicyclooctane, fluorenyl, phenyl,naphthyl, indanyl, adamantyl, or tetrahydronaphthyl (tetralin).Moreover, the ring structures described herein may be attached to one ormore indicated pendant groups via any carbon atom which results in astable structure. The term “substituted” as used in conjunction withcarbocyclic ring structures means that hydrogen atoms attached to thering carbon atoms of ring structures described herein may be substitutedby one or more of the substituents indicated for that structure if suchsubstitution(s) would result in a stable compound.

The term “aryl” which is included with the term “carbocyclic ringstructure” refers to an unsubstituted or substituted aromatic ring,substituted with one, two or three substituents selected from loweralkoxy, lower alkyl, lower alkylamino, hydroxy, halogen, cyano,hydroxyl, mercapto, nitro, thioalkoxy, carboxaldehyde, carboxyl,carboalkoxy and carboxamide, including but not limited to carbocyclicaryl, heterocyclic aryl, and biaryl groups and the like, all of whichmay be optionally substituted. Preferred aryl groups include phenyl,halophenyl, loweralkylphenyl, napthyl, biphenyl, phenanthrenyl andnaphthacenyl.

The term “arylalkyl” which is included with the term “carbocyclic aryl”refers to one, two, or three aryl groups having the number of carbonatoms designated, appended to an alkyl group having the number of carbonatoms designated. Suitable arylalkyl groups include, but are not limitedto, benzyl, picolyl, naphthylmethyl, phenethyl, benzyhydryl, trityl, andthe like, all of which may be optionally substituted.

The term “phenyl” as used herein refers to a six carbon containingaromatic ring which can be variously mono- or poly-substituted with H,C₁–C₆ alkyl, hydroxyl, C₁–C₆ alkoxy, amino, mono-C₁–C₆ alkylamino,di-C₁–C₆ alkylamino, nitro, fluoro, chloro, bromo, iodo,hydroxycarbonyl, or C₁–C₆ alkoxycarbonyl.

As used herein, the term “heterocyclic ring” or “heterocyclic ringsystem” is intended to mean a substituted or unsubstituted memberselected from the group consisting of a stable monocyclic ring havingfrom 5–7 members in the ring itself and having from 1 to 4 hetero ringatoms selected from the group consisting of N, O and S; a stablebicyclic ring structure having a total of from 7 to 12 atoms in the tworings wherein at least one of the two rings has from 1 to 4 hetero atomsselected from N, O and S, including bicyclic ring structures wherein anyof the described stable monocyclic heterocyclic rings is fused to ahexane or benzene ring; and a stable tricyclic heterocyclic ringstructure having a total of from 10 to 16 atoms in the three ringswherein at least one of the three rings has from 1 to 4 hetero atomsselected from the group consisting of N, O and S. Any nitrogen andsulfur atoms present in a heterocyclic ring of such a heterocyclic ringstructure may be oxidized. Unless indicated otherwise the terms“heterocyclic ring” or “heterocyclic ring system” include aromaticrings, as well as non-aromatic rings which can be saturated, partiallysaturated or fully saturated non-aromatic rings. Also, unless indicatedotherwise the term “heterocyclic ring system” includes ring structureswherein all of the rings contain at least one hetero atom as well asstructures having less than all of the rings in the ring structurecontaining at least one hetero atom, for example bicyclic ringstructures wherein one ring is a benzene ring and one of the rings hasone or more hetero atoms are included within the term “heterocyclic ringsystems” as well as bicyclic ring structures wherein each of the tworings has at least one hetero atom. Moreover, the ring structuresdescribed herein may be attached to one or more indicated pendant groupsvia any hetero atom or carbon atom which results in a stable structure.Further, the term “substituted” means that one or more of the hydrogenatoms on the ring carbon atom(s) or nitrogen atom(s) of the each of therings in the ring structures described herein may be replaced by one ormore of the indicated substituents if such replacement(s) would resultin a stable compound. Nitrogen atoms in a ring structure may bequaternized, but such compounds are specifically indicated or areincluded within the term “a pharmaceutically acceptable salt” for aparticular compound. When the total number of O and S atoms in a singleheterocyclic ring is greater than 1, it is preferred that such atoms notbe adjacent to one another. Preferably, there are no more that 1 O or Sring atoms in the same ring of a given heterocyclic ring structure.

Examples of monocylic and bicyclic heterocylic ring systems, inalphabetical order, are acridinyl, azocinyl, benzimidazolyl,benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,benzisothiazolyl, benzimidazalinyl, carbazolyl, 4aH-carbazolyl,carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl,furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl,indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl,isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl(benzimidazolyl), isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl,octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl,oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl,pyroazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pryidooxazole,pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,6H-1,2,5-thiadazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl,thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl,triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,1,3,4-triazolyl and xanthenyl. Preferred heterocyclic ring structuresinclude, but are not limited to, pyridinyl, furanyl, thienyl, pyrrolyl,pyrazolyl, pyrrolidinyl, imidazolyl, indolyl, benzimidazolyl,1H-indazolyl, oxazolinyl, or isatinoyl. Also included are fused ring andspiro compounds containing, for example, the above heterocylic ringstructures.

As used herein the term “aromatic heterocyclic ring system” hasessentially the same definition as for the monocyclic and bicyclic ringsystems except that at least one ring of the ring system is an aromaticheterocyclic ring or the bicyclic ring has an aromatic or non-aromaticheterocyclic ring fused to an aromatic carbocyclic ring structure.

The terms “halo” or “halogen” as used herein refer to Cl, Br, F or Isubstituents. The term “haloalkyl”, and the like, refer to an aliphaticcarbon radicals having at least one hydrogen atom replaced by a Cl, Br,F or I atom, including mixtures of different halo atoms. Trihaloalkylincludes trifluoromethyl and the like as preferred radicals, forexample.

The term “methylene” refers to —CH₂—.

The term “pharmaceutically acceptable salts” includes salts of compoundsderived from the combination of a compound and an organic or inorganicacid. These compounds are useful in both free base and salt form. Inpractice, the use of the salt form amounts to use of the base form; bothacid and base addition salts are within the scope of the presentinvention.

“Pharmaceutically acceptable acid addition salt” refers to saltsretaining the biological effectiveness and properties of the free basesand which are not biologically or otherwise undesirable, formed withinorganic acids such as hydrochloric acid, hydrobromic acid, sulfuricacid, nitric acid, phosphoric acid and the like, and organic acids suchas acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalicacid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaricacid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicyclic acid and the like.

“Pharmaceutically acceptable base addition salts” include those derivedfrom inorganic bases such as sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum salts andthe like. Particularly preferred are the ammonium, potassium, sodium,calcium and magnesium salts. Salts derived from pharmaceuticallyacceptable organic nontoxic bases include salts of primary, secondary,and tertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperizine, piperidine,N-ethylpiperidine, polyamine resins and the like. Particularly preferredorganic nontoxic bases are isopropylamine, diethylamine, ethanolamine,trimethamine, dicyclohexylamine, choline, and caffeine.

“Biological property” for the purposes herein means an in vivo effectoror antigenic function or activity that is directly or indirectlyperformed by a compound of this invention that are often shown by invitro assays. Effector functions include receptor or ligand binding, anyenzyme activity or enzyme modulatory activity, any carrier bindingactivity, any hormonal activity, any activity in promoting or inhibitingadhesion of cells to an extracellular matrix or cell surface molecules,or any structural role. Antigenic functions include possession of anepitope or antigenic site that is capable of reacting with antibodiesraised against it.

In the compounds of this invention, carbon atoms bonded to fournon-identical substituents are asymmetric. Accordingly, the compoundsmay exist as diastereoisomers, enantiomers or mixtures thereof. Thesyntheses described herein may employ racemates, enantiomers ordiastereomers as starting materials or intermediates. Diastereomericproducts resulting from such syntheses may be separated bychromatographic or crystallization methods, or by other methods known inthe art. Likewise, enantiomeric product mixtures may be separated usingthe same techniques or by other methods known in the art. Each of theasymmetric carbon atoms, when present in the compounds of thisinvention, may be in one of two configurations (R or S) and both arewithin the scope of the present invention.

COMPOUND EMBODIMENTS OF THE INVENTION

Compounds of formula (I), formula (II), formula (III), formula (IV),formula (V), formula (VI), formula (VII) and formula (VIII) belowrepresent one embodiment of the invention:

-   A is selected from the group consisting of aryl, substituted aryl,    heteroaryl, substituted heteroaryl, alkylaryl, and alkylheteroaryl.-   W is selected from the group consisting of aryl, substituted aryl,    heteroaryl, and substituted heteroaryl.-   E is selected from the group consisting of H, —C₁–C₈ alkyl,    polyhaloalkyl, —C₃₋₈cycloalkyl, aryl, alkylaryl, substituted aryl,    heteroaryl, and substituted heteroaryl.-   D is selected from the group consisting of NR¹—(C═O)—R², —O—R¹;

wherein:

-   R¹ is independently selected from the group consisting of:    -   H, C₁–C₈ alkyl, polyhaloalkyl, —C₃₋₈-cycloalkyl, aryl,        alkylaryl, substituted aryl, heteroaryl, substituted heteroaryl,        —(C═O)—C₁–C₈ alkyl, —(C═O)-aryl, —(C═O)-substituted aryl,        —(C═O)-heteroaryl and —(C═O)-substituted heteroaryl;-   R² is selected from the group consisting of: aryl, substituted aryl,    heteroaryl, substituted heteroaryl, or-   R¹ and R² can be direct linked or can be indirectly linked through a    carbon chain that is from 1 to about 8 carbon atoms in length,-   n is 0–4,-   m is 0 or 1,-   y is 0–4 and-   Q is independently C or N, with the proviso that when Q is a ring    carbon atom, each ring carbon atom is independently substituted by    X, wherein-   X is in each case a member independently selected from the group    consisting of:    -   H, halogen, polyhaloalkyl, —OR³, —SR³, —CN, —NO₂, —SO₂R³,        —C₁₋₁₀-alkyl, —C₃₋₈-cycloalkyl, aryl, aryl-substituted by 1–4 R³        groups, amino, amino-C₁₋₈-alkyl, C₁₋₃-acylamino,        C₁₋₃-acylamino-C₁₋₈-alkyl, C₁₋₆-alkylamino, C₁₋₆-alkylamino C₁₋₈        alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl, C₁₋₆        alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl, carboxy-C₁₋₆-alkyl,        C₁₋₃-alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl, carboxy        C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl, and a 5 to 10        membered fused or non-fused aromatic or nonaromatic heterocyclic        ring system, having 1 to 4 heteroatoms independently selected        from N, O, and S, with the proviso that the carbon and nitrogen        atoms, when present in the heterocyclic ring system, are        unsubstituted, mono- or disubstituted independently with 0–2 R⁴        groups, and        wherein R and R⁴ are each independently selected from the group        consisting of:    -   H, halogen, —CN, —NO₂, —C₁₋₁₀ alkyl, C₃₋₈-cycloalkyl, aryl,        amino, amino-C₁₋₈-alkyl, C₁₋₃-acylamino,        C₁₋₃-acylamino-C₁₋₈-alkyl, C₁₋₆-alkylamino, C₁₋₆-alkylamino C₁₋₈        alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl, C₁₋₆        alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl, carboxy-C₁₋₆-alkyl,        C₁₋₃-alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl,        carboxy-C₁₋₆-alkyloxy, hydroxy, hydroxy-C₁₋₆-alkyl, -thio and        thio-C₁₋₆-alkyl.-   Y is selected from the group consisting of O, S, N—OR⁵, and NR⁵,    wherein R⁵ is selected from the group consisting of:    -   H, C₁₋₁₀ alkyl, C₃₋₈-cycloalkyl, and CN; and-   Z is selected from the group consisting of NR¹ and O.

The invention also covers all pharmaceutically acceptable salts andprodrugs of the compounds of formula I to formula VIII.

In a preferred embodiment of the invention, compounds of formulae(I)–(VIII)

-   A is selected from the group consisting of:

-   Y is selected from the group consisting of O, S, N—OR⁵ and NR⁵.-   E is selected from the group consisting of H, or C₁₋₈ alkyl.-   W is selected from the group consisting of:

-   D is selected from the group consisting of:

In a more preferred embodiment of the invention, compounds of formulae(I) to (VIII) include the compounds wherein:

-   -   D is selected from the group consisting of:

In another preferred embodiment of the invention, compounds of formulae(I)(VI) include the compounds set forth below in Tables 1–4:

TABLE 1 Formula Ia

R₂ R₁ W Y A

H

O

H

O

H

S

H

N—C≡N

H

O

H

NH

Me

NH

N—C≡N

Me

O

H

O

H

TABLE 2 Formula Ib

X W Y A 3-Br

O

3-Cl

NH

4-OMe

O

H

N—C≡N

3,4-diMe

NH

3-SO₂Me

O

TABLE 3 A. Formula Ic

Y A O

NH

O

N—C≡N

NH

O

TABLE 4 B. Formula Id

R₁ R₂ W H

Me

H

H

Me

H

Examples of specific preferred compounds are listed below:

Preparation of Compounds of the Invention

A compound of formulae (I)–(VIII) may be prepared by various methods asoutlined in the following documents: J. Med. Chem., 33, 23–93–2407(1990); Biorg. & Med. Chem. Letts., Vol. 2, No. 9, pp. 987–992 (1992);J. Med. Chem., 35, 3012–3016 (1992); U.S. Pat. No. 5,234,955 (1993),U.S. Pat. No. 5,354,778 (1994); U.S. Pat. No. 5,565,494 (1996); U.S.Pat. No. 5,594,028 (1997); U.S. Pat. No. 5,302,724 (1994); and WO97/08145, which are incorporated herein in their entirety by reference.Other well-known heterocyclic and carbocyclic synthetic procedures aswell as modification of the procedures that are incorporated above maybe utilized.

Compounds of formulae (I)–(VIII) may be isolated using typical isolationand purification techniques known in the art, including, for example,chromatographic and recrystallization methods.

In compounds of formula formulae (I)–(VIII) of the invention, carbonatoms to which four non-identical substituents are bonded areasymmetric. Accordingly, a compound of formulae (I)–(VIII) may exist asenantiomers, diastereomers or a mixture thereof. The enantiomers anddiastereomers may be separated by chromatographic or crystallizationmethods, or by other methods known in the art. The asymmetric carbonatom when present in a compound of formulae (I)–(VIII) of the invention,may be in one of two configurations (R or S) and both are within thescope of the invention. The presence of small amounts of the opposingenantiomer or diastereomer in the final purified product does not affectthe therapeutic or diagnostic application of such compounds.

According to the invention, compounds of formulae (I)–(VIII) may befurther treated to form pharmaceutically acceptable salts. Treatment ofa compound of the invention with an acid or base may form, respectively,a pharmaceutically acceptable acid addition salt and a pharmaceuticallyacceptable base addition salt, each as defined above. Various inorganicand organic acids and bases known in the art including those definedherein may be used to effect the conversion to the salt.

The invention also relates to pharmaceutically acceptable isomers,hydrates, and solvates of compounds of formulae (I)–(VIII). Compounds offormulae (I)–(VIII) may also exist in various isomeric and tautomericforms including pharmaceutically acceptable salts, hydrates and solvatesof such isomers and tautomers.

This invention also encompasses prodrug derivatives of the compounds offormulae (I)–(VIII). The term “prodrug” refers to a pharmacologicallyinactive derivative of a parent drug molecule that requiresbiotransformation, either spontaneous or enzymatic, within the organismto release the active drug. Prodrugs are variations or derivatives ofthe compounds of formulae (I)–(VIII) of this invention which have groupscleavable under metabolic conditions. Prodrugs become the compounds ofthe invention which are pharmaceutically active in vivo when theyundergo solvolysis under physiological conditions or undergo enzymaticdegradation. Prodrug compounds of this invention may be called single,double, triple, etc., depending on the number of biotransformation stepsrequired to release the active drug within the organism, and indicatingthe number of functionalities present in a precursor-type form. Prodrugforms often offer advantages of solubility, tissue compatibility, ordelayed release in the mammalian organism (Bundgard, Design of Prodrugs,pp. 7–9, 21–24, Elsevier, Amsterdam (1985); Silverman, The OrganicChemistry of Drug Design and Drug Action, pp. 352–401, Academic Press,San Diego, Calif. (1992)). Prodrugs commonly known in the art includeacid derivatives well known to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acids with a suitablealcohol, or amides prepared by reaction of the parent acid compound withan amine, or basic groups reacted to form an acylated base derivative.Moreover, the prodrug derivatives of this invention may be combined withother features herein taught to enhance bioavailability.

EXAMPLE 1

Preparation of 5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}(cyanoimino)methyl)amino]-2-methylphenyly}benzo[c]azolidine-1,3-dione

A solution of N-(4-amino-2-methylphenyl)-4-chlorophthalimide (0.14 g,0.5 mmol) and dimethyl N-cyanodithioiminocarbonate (0.13 g, 1 mmol) inpyridine (1.3 mL) was stirred at 115° C. for 8 hr. The reaction mixturewas then cooled and concentrated in vacuo. To a solution of this crudeintermediate (56 mg, 0.11 mol) in pyridine (0.7 mL) was added DBU (33μL, 0.22 mmol) and 5-chlorothiophene-2-sulfonamide (44 mg, 0.22 mmol).The reaction mixture was heated at 115° C. for 23 hr with addition ofDMAP (10 mg) after 2 hr. Acidification and HPLC purification yielded(2Z)-2-aza-3-{[(5-chloro(2-thienyl))sulfonyl]amino}-3-{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}prop-2-enenitrile(14 mg, 24%). ES-MS (M+H)+=534, 536 (Cl).

¹H-NMR (DMSO-d₆): δ 2.03 (s, 3H), 7.06–7.07 (d, 1H), 7.18–7.20 (d, 1H),7.30–7.31 (d, 1H), 7.37 (s, 2H), 7.93–7.94 (2H), 8.03 (d, 1H), 8.84 (s,1H).

EXAMPLE 1061

Preparation of(5-chloro(2-thienyl))-N-({[4-(5-chloro-1,3-dioxobenzo[c]azolin-2-yl)-3-methylphenyl]amino}thioxomethyl)carboxamideA. Synthesis of 5-chlorothiophene-2-carbonyl chloride

To a chilled solution of 5-chlorothiophene-2-carboxylic acid (0.16 g,1.0 mmol) in EtOAc (3 mL) and DMF (1 drop) was added neat oxalylchloride (92 μL, 1.05 mmol). The reaction mixture was stirred cold for 2hr and concentrated in vacuo to give crude 5-chlorothiophene-2-carbonylchloride.

B. Synthesis of(5-chloro(2-thienyl))-N-({[4-(5-chloro-1,3-dioxobenzor[c]azolin-2-yl)-3-methylphenyl]amino}thioxomethyl)carboxamide

To a suspension of KSCN (29 mg, 0.3 mmol) in dry acetonitrile (0.2 mL)was added a solution of the crude acid chloride (36 mg, 0.2 mmol) inCH₃CN (0.2 mL). The resulting suspension was stirred at room temp for 30min. This acylthioisocyanate in situ was added to a suspension ofN-(4-amino-2-methylphenyl)-4-chlorophthalimide (58 mg, 0.2 mmol) inCH₃CN. The reaction mixture was stirred at room temp for 1 hr, filteredand dried to give pure(5-chloro-(2-thienyl))-N-({[4-(5-chloro-1,3-dioxobenzo[c]azolin-2-yl)-3-methylphenyl]amino}thioxomethyl)carboxaimide(66 mg, 70%). ES-MS (M+H)+=490, 492 (Cl).

EXAMPLE 1062

Preparation of5-chloro-2-(4-{[5-(5-chloro(2-thienyl))(4H-1,2,4-triazol-3-yl)]amino}-2-methylphenyl)benzo[c]azoline-1,3-dione

To a suspension of(5-chloro(2-thienyl))-N-({[4-(5-chloro-1,3-dioxobenzo[c]azolin-2-yl)-3-methylphenyl]amino}thioxomethyl)carboxamide(15 mg, 0.030 mmol) and hydrazine dihydrochloride (4 mg, 0.038 mmol) inDMF (0.3 mL) was added HgO (7 mg, 0.032 mmol). The reaction was stirredat room temp for 17 hr, and HPLC purified to give the desired product5-chloro-2-(4-{[5-(5-chloro(2-thienyl))(4H-1,2,4-triazol-3-yl)]amino}-2-methylphenyl)benzo[c]azoline-1,3-dione(2 mg) (ES-MS (M+H)+=470, 472) and the aminoguanidine intermediateN-((1E)-2-aza-1-{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-2-aminovinyl)(5-chloro(2-thienyl))carboxamide(2 mg) ES-MS (M+H)+=488, 490 (2Cl).

EXAMPLE 1063

Preparation of3-(4-{[5-(5-chloro-2-thienyl)-4H-1,2,4-triazol-3-yl]amino}phenyl)-1,3-dihydroquinazoline-2,4-dionewas executed using the same methodology as shown in Examples 1061 and1062, using 3-(4-aminophenyl)-1,3-dihydroquinazoline-2,4-dione as theaniline in step B from Example 1061. ES-MS (M+H)+=437, 439 (Cl).

EXAMPLE 1064

To a solution of triphosgene (22 mg, 0.074 mmol) in CH₂Cl₂ (1 mL) wasadded a suspension of N-(4-amino-2-methylphenyl)-4-chlorophthalimide (57mg, 0.2 mmol) in CH₂Cl₂ (1.5 mL) and diethylisopropylamine (70 μL)dropwise over 10 min. The reaction mixture was stirred for 10 min, thena suspension of benzenesulfonylhydrazide (52 mg, 0.3 mmol) in CH₂Cl₂(1.5 mL) and DIEA (35 μL) was added. The mixture was stirred at roomtemp for 17 hr, acidified and HPLC purified to giveN-[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl][2-(phenylsulfonyl)hydrazino]-carboxamide(43 mg, 46%). ES-MS (M+H)+=485, 487. ¹H-NMR (DMSO-d₆): δ 2.00 (s, 3H),7.13–7.15 (d, 1H), 7.28–7.31 (d, 1H), 7.34 (s, 1H), 7.55–7.67 (m, 3H),7.82–7.84 (m, 2H), 7.93 (s, 2H), 8.02 (s, 1H).

EXAMPLE 1065

Preparation of{[(5-chloro(2-thienyl))methyl]amino}-N-[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]carboxamideA. Synthesis of5-chloro-2-[(1,3,5,7-tetraazatricyclo[3,3,1,1(3,7)]decyl)methyl]thiophene

To a suspension of hexamethylenetetramine (HMTA) (3.12 g, 22.2 mmol) inCHCl₃ (35 mL) was added 2-chloro-5-chloromethylthiophene (1.02 mL, 8.46mmol). The reaction mixture was heated at reflux for 4 hr, cooled, andfiltered to give white solid5-chloro-2-[(1,3,5,7-tetraazatricyclo-[3.3.1.1 (3,7)]decyl)methyl]thiophene (2.28 g, 88%). ES-MS (M)+=271, 273 (Cl). ¹H-NMR(DMSO-d₆): δ 4.27 (s, 2H), 4.39–4.57 (ABq, 6H), 5.06 (s, 6H), 7.21–7.24(ABq, 2H).

B. Synthesis of (5-chloro-2-thienyl)methylamine

To a solution of5-chloro-2-[(1,3,5,7-tetraazatricyclo[3.3.1.1(3,7)]decyl)methyl]thiophene(2.15 g, 7 mmol) in methanol (10 mL) and water (5 mL) was added conc.HCl (5 mL). The reaction mixture was refluxed for 3 hr, poured ontowater and washed with ethyl ether. The aqueous layer was basified with4N NaOH and extracted into ethyl ether, washed with brine, dried andconcentrated in vacuo to give (5-chloro-2-thienyl)methylamine (0.8 g,78%).

C. Synthesis of{[(5-chloro(2-thienyl))methyl]amino}-N-[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]carboxamide

To a solution of triphosgene (22 mg, 0.074 mmol) in CH₂Cl₂ (1 mL) wasadded a suspension of N-(4-amino-2-methylphenyl)-4-chlorophthalimide (57mg, 0.2 mmol) in CH₂Cl₂ (1.5 mL) and diethylisopropylamine (70 μL)dropwise over 10 min. The reaction mixture was stirred for 10 min, thena solution (5-chloro-2-thienyl)methylamine from step B (47 mg, 0.32mmol) in CH₂Cl₂ (1 mL) and DIEA (35 μL) was added. The mixture wasstirred at room temp for 17 hr, acidified and HPLC purified to give{[(5-chloro(2-thienyl))methyl]amino}-N-[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]carboxamide(18 mg, 20%). ES-MS (M+H)+=460, 462 (Cl). ¹H-NMR (DMSO-d₆): δ 2.02 (s,3H), 4.34–4.36 (d, 2H), 6.77–6.80 (t, 1H), 6.82–6.93 (2d, 2H), 7.14–7.16(d, 1H), 7.29–7.32 (dd, 1H), 7.41 (d, 1H), 7.93 (ABq, 2H), 8.02 (s, 1H),8.77 (s, 1H).

EXAMPLE 1066

Preparation of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}thioxomethyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dione A. Synthesis of4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylbenzenisothiocyanate

To a slurry of 150 mg (0.52 mmol) of2-(4-amino-2-methylphenyl)-5-chlorobenzo[c]azoline-1,3-dione in 2 mL ofacetone, was added 41 uL (0.54 mmol) of thiophosgene. The yellow slurrydissolved whereupon a white precipitate formed. After 1 h, this solidwas collected by filtration and dried to give 127 mg (74%) of thedesired product.

B. Synthesis of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}thioxomethyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dione

To a slurry of 51 mg (0.156 mmol) of the isothiocyanate prepared aboveand 31 mg (0.156 mmol) of 5-chlorothiophenesulfonamide in 300 μL of DMSOwas added 26 μL of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). After a fewminutes the reaction mixture was diluted with 1.2 mL of water andacidified with acetic acid to pH 4 when a precipitate was formed,collected, and dried to give 77 mg (94%) of the titled compound. ES-MS(M+H)+=525.8 (2Cl).

EXAMPLE 1067

Preparation of2-[4-({(1Z)-2-aza-2-[(5-chloro(2-thienyl))sulfonyl]-1-methylthiovinyl}amino)-2-methylphenyl]-5-chlorobenzo[c]azolidine-1,3-dione

To a solution of 20 mg (0.038 mmol)5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}thioxomethyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dionein 780 μL of acetone and 63 μL of 0.6M NaHCO₃ was added 5.9 μL of methyliodide. After 2 h, the reaction mixture was acidified with acetic acidand the precipitate was collected and dried to give 13 mg (63%) of thetitled compound.

EXAMPLE 1068

Preparation of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}iminomethyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dione

A 15 mg portion of the5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}thioxomethyl)-amino]-2-methylphenyl}benzo[c]azolidine-1,3-dionewas dissolved in 120 μL of DMF containing 6.6 mg of conc. ammoniumhydroxide and 6 mg of mercuric oxide was added. After stirring for 18 h,the mercuric sulfide was filtered off and the solution was purified byreversed phase HPLC to give 1 mg (7%) of a white solid. ES-MS (M+H)+=509(2Cl).

EXAMPLE 1069

Preparation of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}(hydroxyimino)methyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dione

The titled compound was prepared in a similar fashion as for Example1068 with a 10% yield after purification. ES-MS (M+H)+=525 (2Cl).

EXAMPLE 1070

Preparation of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}methyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dioneA. Synthesis of2-[4-((1E)-1-aza-2-ethoxyvinyl)-2-methylphenyl]-5-chlorobenzo[c]azolidine-1,3-dione

A 50 mg (0.175 mmol) portion of2-(4-amino-2-methylphenyl)-5-chlorobenzo[c]azoline-1,3-dione in 2 mL oftriethylorthoformate was heated to reflux for 1 h then distilled toleave a solid, 598 mg (100%).

B. Synthesis of5-chloro-2-{4-[({[(5-chloro(2-thienyl))sulfonyl]amino}methyl)amino]-2-methylphenyl}benzo[c]azolidine-1,3-dione

A 100 mg sample of2-[4-((1E)-1-aza-2-ethoxyvinyl)-2-methylphenyl]-5-chlorobenzo[c]-azolidine-1,3-dioneplus 58 mg (0.29 mmol) of 5-chlorothiophenesulfonamide was slurried in1.2 mL of MeOH, heated to reflux for 2 h, and the methanol was distilledoff. The remaining solid was triturated with ACN/MeOH, filtered, andconcentrated to afford 104 mg (87%) of the titled compound. ES-MS(M+H)+=494.

EXAMPLE 1071

Preparation of5-chloro-2-[4-({1-[(5-chloro(2-thienyl))sulfonyl](1,2,3,4-tetraazol-5-yl)}amino)-2-methylphenyl]benzo[c]azolidine-1,3-dione

The titled compound was prepared in a similar fashion as for Example1068 to yield a 18% yield after RP-HPLC purification. ES-MS (M+H)+=535(2Cl).

EXAMPLE 1072

5-chloro-2-[4-({1-[(5-chloro(2-thienyl))sulfonyl](1,2,3,4-tetraazol-5-yl)}amino)-2-methylphenyl]benzo[c]azolidine-1,3-dioneA. Synthesis of chlorophenylsulfoxide

To 2 g (12 mmol) of sodium benzenesulfinic acid was added 5 mL ofthionyl chloride and stirred at 0° C. for 4 h. The product was isolatedby bulb-to-bulb distillation (180° C. @ 4 mmHg) to afford 1.25 g (64%)of the liquid benzenesulfinic chloride.

B. Synthesis of benzenesulfinamide

The chlorophenylsulfoxide (500 mg, 3.2 mmol) was dissolved in 5 mL ofdiethyl ether at 0° C. and anhydrous ammonia was bubbled through untilno more precipitate is formed. The solution was filtered andconcentrated to afford a solid which was recrystallized from water toafford 152 mg (35%) of benzenesulfinamide.

C. Synthesis of5-chloro-2-[4-({1-[(5-chloro(2-thienyl))sulfonyl](1,2,3,4-tetraazol-5-yl)}amino)-2-methylphenyl]benzo[c]azolidine-1,3-dione

A 16 mg portion of benzenesulfinamide and 47 mg of2-(4-amino-2-methylphenyl)-5-chlorobenzo[c]azoline-1,3-dione wasdissolved in 232 uL of CAN followed by 18 μL of DBU. The reaction wasstirred at 23° C. for 1 h and purified by RP-HPLC to give 20 mg (38%) ofthe desired material. ES-MS (M+H)+=454 (2Cl).

EXAMPLE 1073

Preparation of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-t-butylphenylsulfonimidoyl)carboxamideA. Synthesis of (tert-butyl)(phenylsulfonyl)amine

In a similar fashion for the preparation of Example 1072 B, tert-butylamine (5 equivalents) was used to prepare the named compound in 75%yield. ¹H-NMR (DMSO-d₆): δ 7.67 (m,2); 7.45 (m,3) 4.83 (br s,1); 1.39(s,9).

B. Synthesis of (tert-butyl)(phenyliminosulfonyl)amine

To a 50 mg-portion (0.25 mmol) of (tert-butyl)(phenylsulfonyl)amine in 5mL of anhydrous THF was added 58 mg (0.26 mmol) of N-chlorosaccharinunder an argon atmosphere. After a few minutes the reaction mixture wascooled to −78° C., and anhydrous ammonia was bubbled through. Afterwarming to 23° C. the solvent was evaporated, the residue was dissolvedin water, extracted 3 times with EtOAc. The combined organic layers werewashed with 5% NaHCO₃, dried (Na₂SO₄), and concentrated to afford 37 mg(68%) of the titled compound. ES-MS (M+H)+=236.

C. Synthesis of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-t-butylphenylsulfonimidovl)carboxamide

In a similar fashion for preparation of Example 1072C, the titledcompound was prepared in 2% yield after RP-HPLC purification. ES-MS(M+H)+=525.

EXAMPLE 1074

Preparation of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-benzylphenylsulfonimidoyl)carboxamideA. Synthesis of (benzyl)(phenylsulfonyl)amine

In a similar fashion for the preparation of Example 1072 Step B,benzylamine (5 equivalents) was used to prepare the named compound in74% yield. ES-MS (M+H)+=232.1. ¹H-NMR (CDCl₃) δ 7.67 (m,2); 7.51 (m,3);7.28 (m,5); 4.28 (ABX,1); 4.23 (ABX,1); 3.89 (X,1).

B. Synthesis of (benzyl)(phenyliminosulfonyl)amine

In a similar fashion as for the preparation of Example 1072 Step C, thetitled compound was prepared in quantitative yield.

C.{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-benzylphenylsulfonimidoyl)carboxamide

In a similar fashion for preparation of Example 1072 Step C, the titledcompound was prepared in 34% yield after RP-HPLC purification. ES-MS(M+H)+=559.

EXAMPLE 1075

Preparation of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-p-methoxybenzylphenylsulfonimidoyl)carboxamideA. Synthesis of (p-methoxybenzyl)(phenyliminosulfonyl)amine

In a similar fashion for the preparation of Example 1072 Step B,p-methoxybenzylamine (5 equivalents) was used to prepare the namedcompound in 66% yield. ES-MS (M+H)+=262. ¹H-NMR (CDCl₃): δ 7.97 (m, 2);7.49 (m, 3); 7.18 (d, 2); 6.81 (d, 2); 4.20 (m, 2); 3.8 (dd, 1); 3.74(s, 3).

B. Synthesis of (p-methoxybenzyl)(phenyliminosulfonyl)amine

In a similar fashion as for the preparation of Example 1072 Step C, thetitled compound was prepared in quantitative yield. Material used innext step without purification.

C. Synthesis of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(N-benzylphenylsulfonimidoyl)carboxamide

In a similar fashion for preparation of Example 1072 Step C, the titledcompound was prepared in 17% yield after RP-HPLC purification. ES-MS(M+H)+=589.

EXAMPLE 1076

Preparation of {[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]amino}-N-(phenylsulfonimidoyl)carboxamide

To 10 mg (0.016 mmol) of{[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]-amino}-N-(N-p-methoxybenzylphenylsulfonimidoyl)carboxamidedissolved in 380 μL of CAN followed by 96 uL of water was added 70 mg(0.13 mmol) of cerric ammonium nitrate. After 20 m, the reaction wascomplete and purified by RP-HPLC to give 1.8 mg (23%) afterlyophilization. ES-MS (M+H)+=469 (Cl).

EXAMPLE 1077

Preparation ofN-{[4-(1,3-dioxobenzo[c]azolidin-2-yl)phenyl]sulfonyl(phenylamino)carboxamide A. Synthesis of4-(1,3-dioxobenzo[c]azolidin-2-yl)benzenesulfonamide

A solution of 1.0 g (5.8 mmol) of p-aminophenylsulfonamide in 3.6 mL ofpyridine was added 878 μL (6.1 mmol) of phthalic dichoride. Afterheating to 60° C. for 18 h, the solution was poured into 1N HCl, cooledto 0° C., the precipitate was collected by filtration, and dried undervacuum to give 1.58 g (90%) of the titled compound.

A. Synthesis ofN—{[4-(1,3-dioxobenzo[c]azolidin-2-yl)phenyl]sulfonyl}(phenylamino)carboxamide

To a solution of 4-(1,3-dioxobenzo[c]azolidin-2-yl)benzenesulfonamide in660 uL of DMSO was added 60 mg (0.40 mmol) of DBU followed by 36 μl(0.33 mmol) of phenylsulfonylisocyanate. After stirring for 0.5 h, themixture was poured into 1N HCl, cooled, and the precipitate wascollected by filtration and dried under vacuum to give 137 mg (100%) ofthe titled compound.

EXAMPLE 1078

Preparation ofN-[(5-chloro(2-thienyl))sulfonyl]-N′-[4-(5-chloro-1,3-dioxobenzo[c]azolidin-2-yl)-3-methylphenyl]ethane-1,2-diamide

A 200 mg (1.0 mmol) of 5-chlorothiophenesulfonamide was slurried in 0.5mL of oxalyl chloride and refluxed for 6 h. The solvent was removed invacuo and 36 mg of the resulting solid was dissolved in 240 μLacetonitrile and treated with 40 mg(0.14 mmol)2-(4-amino-2-methylphenyl)-5-chlorobenzo[c]azoline-1,3-dione. Afterstirring for 1 h, the solvent was removed and the residue was purifiedby RP-HPLC to give 33 mg (44%) of the titled compound. ES-MS(M+H)+=538(2Cl).

EXAMPLE 1079

Preparation of[4-(1,3-dioxobenzo[c]azolidin-2-yl)phenyl]-N-[(5-chloro(2-thienyl))sulfonyl]-carboxamideA. Synthesis ofN-[(5-chloro(2-thienyl))sulfonyl](4-nitrophenyl)carboxamide

A 85 mg portion (0.43 mmol) of 5-chlorothiophenesulfonamide dissolved in2 mL of acetone was treated with 110 μL of 4N NaOH (0.43 mmol) followedby addition of 80 mg (0.43 mmol) of 4-nitrobenzoyl chloride. Afterstirring for 12 h, the solution was acidfied 1N HCl and the precipitatewas collected by filtration and dried. Recrystallization fromEtOAc/hexane afforded 120 mg (81%) of the titled compound. ES-MS(M+H)+=347 (Cl).

B. Synthesis of(4-aminophenyl)-N-[(5-chloro(2-thienyl))sulfonyl]carboxamide

A 74 mg portion (0.21 mmol) of[4-(1,3-dioxobenzo[c]azolidin-2-yl)phenyl]-N-[(5-chloro(2-thienyl))sulfonyl]-carboxamide,192 mg (0.84 mmol) of tin dichloride dihydrate were combined anddissolved in 1.4 mL of EtOAc. The mixture was refluxed for 4 h, filteredthrough celite, and concentrated in vacuo to afford a solid which waspurified on silica gel eluting with 10% MeOH/CH₂Cl₂ to give aquantitative yield of the titled compound. ES-MS (M+H)+=317 (Cl).

C. Synthesis of[4-(1,3-dioxobenzo[c]azolidin-2-yl)phenyl]-N-[(5-chloro(2-thienyl))sulfonyl]carboxamide

A 22 mg portion (0.070) of(4-aminophenyl)-N-[(5-chloro(2-thienyl))sulfonyl]carboxamide wascombined with 15 mg (0.10 mmol) of phthalic anhydride in 140 μL of DMF.After 18 h of heating at 110° C., the mixture was cooled and purified byRP-HPLC, to give 20 mg (55%) of the desired compound. ES-MS (M+H)⁺=447(Cl).

EXAMPLE 1080

[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]-N-[(5-chloro(2-thienyl))sulfonyl]carboxamide

A 44 mg portion (0.14) of(4-aminophenyl)-N-[(5-chloro(2-thienyl))sulfonyl]carboxamide wascombined with 25 mg (0.14 mmol) of methyl 2 isocyanatobenzoate in 500 uLof THF followed by the addition of 1 equivalent of TEA (24 μL) and 1equivalent of DBU. After 18 h, the mixture was purified by RP-HPLC, togive 21 mg (34%) of the desired compound. ES-MS (M+H)⁺=462 (Cl).

General Procedure for Synthesizing Phthalimide Targets

A. General Procedure for phthalic anhydride reaction

A mixture of phthalic anhydride (0.96 g, 6.5 mmol) and the substitutednitroaniline (5 mmol) in DMF (10 mL) was heated to 120° C. for 22 hr.The reaction was then concentrated and chromatographed on silica gelusing CH₂Cl₂/hexane mixtures as eluent to give pure product in 35–65%yields, depending on the substituents.

B-1. General Reduction Procedure 1

A suspension of the nitrophenyl intermediate (0.75 mmol) and tin(II)chloride dihydrate (3 mmol) in ethyl acetate (4.5 mL) was heated at 70°C. for 2 hr. The reaction mixture was then poured onto ice (25 mL),basified with 5% NaHCO₃ (13 mL), extracted with EtOAc, washed withbrine, dried and concentrated in vacuo to give pure aniline product invery good yields, typically >90%.

B-2. General Reduction Procedure 2

To a suspension of the nitrophenyl intermediate (0.8 mmol) in methanol(3 mL), ethyl acetate (2 mL) and 2 N HCl (0.4 mL, 0.8 mmol) was added10% Pd/C (43 mg, 0.04 mmol) under argon. The reaction mixture washydrogenated under 1 atm H₂ for 2 hr, filtered through Celite andconcentrated in vacuo to give the aniline hydrochloride salt in verygood yields.

C. General Procedure for Coupling the Aniline Intermediate with5-chlorothiophenesulfonamides to Form Sulfonyl Ureas

To a suspension of 5-chlorothiophene-2-sulfonamide (40 mg, 0.2 mmol) andDSC (61 mg, 0.24 mmol) in dry acetonitrile (1 mL) was added DBU (60 μL,0.4 mmol). The resulting solution was stirred at room temp for 16 hr.The aniline from step B (0.2 mmol) was then added as a solid withadditional acetonitrile (1 mL), and the reaction was heated to 70° C.and stirred for another 17 hr. Acidification and HPLC purification ofthe crude reaction product gave the final compounds in varying yields(20–70%).

EXAMPLE 7 AND EXAMPLES 1081–1093

The targets above were prepared using the procedures outlined in Scheme1, steps A–C using a variety of substituted nitroanilines, where R=3-Cl;3-CN; 3-CF₃; 3-F; 3-Br; 3-OMe; 3-iPr; 2-CF₃; 2-Cl, 5-Me; 2-NMe₂, 5-Cl;2,5-diMe; 3,5-diMe; 3,5-diCl; 2-OMe, 5-Me; and X═N, R═H.

EXAMPLE 7N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-chlorophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=496, 498 (2Cl). EXAMPLE 1081N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-cyanophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=487, 489 (Cl). EXAMPLE 1082N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-(trifluoromethyl)phenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide. ES-MS(M+H)+=530, 532 (Cl). EXAMPLE 1083N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-fluorophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=530, 532 (Cl). EXAMPLE 1084N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-bromophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=540, 542 (Cl, Br). EXAMPLE 1085N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-methoxyphenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=492.0 (Cl). EXAMPLE 1086N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3-(methylethyl)phenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=490.9 (Cl). EXAMPLE 1087N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-2-(trifluoromethyl)phenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide. ES-MS(M+H)+=463, 465 (Cl). EXAMPLE 1088N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-2-chloro-5-methylphenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=510, 512 (2Cl). EXAMPLE 1089N-[2-(dimethylamino)-4-(1,3-dioxobenzo[c]azolidin-2-yl)-5-chlorophenyl]{[(5-chloro(2-thienyl))-sulfonyl]amino}carboxamide.ES-MS (+H)+=539, 541 (2Cl). EXAMPLE 1090N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-2,5-dimethylphenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=490.0 (Cl). EXAMPLE 1091N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3,5-dimethylphenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=490.0 (Cl). EXAMPLE 1092N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-3,5-dichlorophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide.ES-MS (M+H)+=529.9, 532.0 (2Cl) EXAMPLE 1093 N-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-5-methyl-2-methoxyphenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=506, 507 (Cl). EXAMPLE 1094

Preparation ofN-[6-(1,3-dioxobenzo[c]azolidin-2-yl)(3-pyridyl)]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamidewas accomplished using the procedures outlined in Scheme 1, steps A–Cusing 2-amino-5-nitropyridine. ES-MS (M+H)+=463, 465 (Cl).

EXAMPLE 1095

Preparation ofN-[6-(1,3-dioxobenzo[c]azolidin-2-yl)(3-phenyl)]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamidewas accomplished using the procedures outlined in Scheme 1, steps A–C.ES-MS (M+H)+=462.0 (Cl).

EXAMPLE 1096

Preparation ofN-[4-(1,3-dioxobenzo[c]azolidin-2-yl)-4-napthyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamidewas accomplished using the procedures outlined in Scheme 1, steps A–C.ES-MS (M+H)+=511.9 (Cl).

EXAMPLE 1097

Preparation ofN-[5-(1,3-dioxobenzo[c]azolidin-2-yl)(2-pyridyl)]{[(5-chloro(2-thienyl))-sulfonyl]amino}carboxamideA. Synthesis of (tert-butoxy)-N-(5-nitro(2-pyridyl)carboxamide

To a solution of 2-amino-5-nitropyridine (0.555 g, 4 mmol) in THF (10mL) was added 1 M NaHMDS in THF (8 mL, 8 mmol). The resulting dark redsuspension was stirred for 15 min, followed by addition of a solution ofBoc anhydride (0.87 mL, 3.8 mmol) in THF (5 mL). The reaction mixturewas stirred at room temp for 21 hr, dilute with EtOAc, washed with 1 NHCl and brine, dried and concentrated in vacuo to give(tert-butoxy)-N-(5-nitro(2-pyridyl))carboxamide (0.63 g, 70%). ES-MS(M+H-tBu)+=184.

B. Synthesis of N-(5-amino(2-pyridyl))(tert-butoxy)carboxamide

To a suspension of (tert-butoxy)-N-(5-nitro(2-pyridyl))carboxamide (0.27g, 1.13 mmol) in methanol (2 mL), ethyl acetate (4 mL) and TEA (0.16 mL)was added 10% Pd/C (60 mg, 0.056 mmol) under argon. The reaction mixturewas hydrogenated under 1 atm H₂ for 20 hr, filtered through Celite andconcentrated in vacuo to giveN-(5-amino(2-pyridyl))(tert-butoxy)carboxamide (0.226 g, 97%). ¹H-NMR(DMSO-d₆): δ 1.40 (s, 9H), 4.92 (br s, 2H), 6.89–6.91 (dd, 1H),7.35–7.37 (d, 1H), 7.58 (d, 1H), 9.06 (s, 1H).

C. Synthesis ofN-[5-(1,3-dioxobenzo[c]azolidin-2-yl)(2-pyridyl)]{[(5-chloro(2-thienyl))-sulfonyl]amino}carboxamide

N-[5-(1,3-dioxobenzo[c]azolidin-2-yl)(2-pyridyl)]{[(5-chloro(2-thienyl))-sulfonyl]amino}carboxamidewas prepared by following the procedure in Scheme 1 step A, followed byTFA deprotection, followed by the coupling procedure outlines in Scheme1 step C. ES-MS (M+H)+=463, 465 (Cl).

EXAMPLE 1098

Preparation ofN-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

A. Synthesis of N-(2-aminopyrimidin-5-yl)(tert-butoxy)carboxamide

To a suspension of 2-amino-5-nitropyrimidine (0.25 g, 1.78 mmol) inmethanol (4 mL) was added tert-butyl (tert-butoxycarbonyloxy)formate(0.5 mL, 2.18 mmol) and 10% Pd/C (96 mg, 0.090 mmol) under argon. Thereaction mixture was hydrogenated under 1 atm H₂ for 5 hr, filteredthrough Celite and concentrated in vacuo to give crudeN-(2-aminopyrimidin-5-yl)(tert-butoxy)carboxamide (0.435 g). ES-MS(M+H)+=211.

B. Synthesis of (tert-butoxy)-N-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]carboxamide

To a solution of N-(2-aminopyrimidin-5-yl)(tert-butoxy)carboxamide(0.237 g, 1.0 mmol) in pyridine (1 mL) was added phthaloyl dichloride(0.144 mL, 1.0 mmol). The resulting suspension was stirred at 45° C. for2 hr, diluted with EtOAc, washed with water and brine, dried andconcentrated in vacuo to give crude(tert-butoxy)-N-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]carboxamide(0.31 g). ES-MS (M+H)+=341; (M+H-tBu)+=285.

C. Synthesis ofN-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

TFA deprotection of(tert-butoxy)-N-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]carboxamideand coupling with 5-chlorothiophenesulfonamide (see Scheme 1 step C)gaveN-[2-(1,3-dioxobenzo[c]azolidin-2-yl)pyrimidin-5-yl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamidein 27% yield. ES-MS (M+H)+=464, 466 (Cl).

EXAMPLE 1099

Preparation ofN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-benzamideA. Synthesis of N-(2-methyl-4-nitrophenyl)benzamide

To a suspension of 2-methyl-4-nitroaniline (0.76 g, 5 mmol) in toluene(25 μL) was added neat benzoyl chloride (0.59 mL, 5.08 mmol). Thereaction mixture was refluxed for 16 hr, cooled and filtered to giveN-(2-methyl-4-nitrophenyl)benzamide (1.21 g, 95%) as a beige solid.ES-MS (M+H)+=257.

B. Synthesis of N-(4-amino-2-methylphenyl)benzamide

A suspension of N-(2-methyl-4-nitrophenyl)benzamide (0.256 g, 1.0 mmol)and tin(II) chloride dihydrate (0.89 g, 3.96 mmol) in ethyl acetate (6mL) was heated at 70° C. for 19 hr. The reaction mixture was thenchilled, poured onto 50 mL ice, basified with 5% NaHCO₃ (20 mL),extracted into EtOAc, washed with brine, dried and concentrated in vacuoto give N-(4-amino-2-methylphenyl)-benzamide (0.22 g, 97%). ES-MS(M+H)+=227, (M+Na)+=249.

C. Synthesis ofN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]benzamide

A solution of 5-chlorothiophene-2-sulfonamide (30 mg, 0.15 mmol) and DSC(46 mg, 0.18 mmol) in CH₃CN (1 mL) and DBU (45 μL) was heated at 40° C.for 1 hr. To this mixture was added N-(4-amino-2-methylphenyl)benzamide(34 mg, 0.15 mmol) with further heating for 3 days. Acidification andHPLC purification gaveN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonyl-amino)-2-methylphenyl]benzamide(24 mg, 35%). ES-MS (M+H)+=450, 452 (Cl).

EXAMPLE 1100

Preparation ofN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-N-methylbenzamideA. Synthesis of N-methyl-N-(2-methyl-4-nitrophenyl)benzamide

To a solution of N-(2-methyl-4-nitrophenyl)benzamide (Example 1099A)(0.38 g, 1.48 mmol) in DMF (2 mL) was added cesium carbonate (1.2 g,3.68 mmol) followed by methyl iodide (0.12 mL, 1.9 mmol). The reactionmixture was stirred at room temp for 2 hr, extracted with EtOAc, washedwith water and brine, dried and concentrated in vacuo to giveN-methyl-N-(2-methyl-4-nitrophenyl)-benzamide (0.38 g, 95%). ES-MS(M+H)+271.

B. Synthesis ofN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-N-methylbenzamide

N-methyl-N-(2-methyl-4-nitrophenyl)benzamide was reduced and coupledwith 5-chloro-thiophenesulfonamide using the same procedure as shown inExample 1099, steps B and C to giveN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-N-methylbenzamide(44 mg, 31%). ES-MS (M+H)+=464, 466 (Cl).

EXAMPLE 1101

Preparation ofN-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](3-chlorophenyl)-carboxamidewas prepared using a similar procedure as that shown in Example 1099steps A–C, using 3-chlorobenzoyl chloride in the first step, instead ofbenzoyl chloride. The final product was obtained in 43% yield. ES-MS(M+H)+=484, 486 (2Cl).

EXAMPLE 1102

Preparation ofN-[2-bromo-4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]benzamide was prepared using a similar procedure as that shown inExample 1099 steps A–C, using 2-bromo-4-nitro-aniline in the first step,instead of 2-methyl-4-nitroaniline. The final product was obtained in64% yield. ES-MS (M+H)+=514, 516, 518 (Cl, Br).

EXAMPLE 1103

Preparation of N-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide A. Synthesis ofN-{4-[(tert-butoxy)carbonylamino]-3-methylphenyl}{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

A solution of 5-chlorothiophene-2-sulfonamide (0.2 g, 1.0 mmol) and DSC(0.307 g, 1.2 mmol) in CH₂Cl₂ (5 mL) and DBU (0.3 mL) was stirred atroom temp for 16 hr. To this mixture was addedN-(4-amino-2-methylphenyl)(tert-butoxy)carboxamide (0.26 g, 1 mmol),followed by heating at 40° C. for 2 hr. Acidification and HPLCpurification gaveN-{4-[(tert-butoxy)carbonylamino]-3-methyl-phenyl}{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide(0.28 g, 63%). ES-MS (M+Na)⁺=468, (M+H−tBu)=390, 392 (Cl).

B. Synthesis of N-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

To a chilled solution ofN-{4-[(tert-butoxy)carbonylamino]-3-methylphenyl}{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide(0.246 g, 0.55 mmol) in CH₂Cl₂ (5 mL) was added neat TFA (1.1 mL). Thereaction mixture was stirred cold for 1.5 hr, concentrated in vacuo,azeotroped with heptane and dried to give N-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide (0.26 g) as the monoTFA salt. ES-MS (M+H)+=346.

EXAMPLES 1104–1116

The compounds above were prepared according to Scheme 4 using thefollowing general synthetic procedure: 1.1 equivalent of the benzoicacid and 1 equivalent of N-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide were dissolved in DMF(0.5 M) and 1.2 equivalent of PyBOP was added. After 2 h, the reactionmixture was directly purified by RP-HPLC to give the targets above whereR=p-CH₃, p-OCH₃, p-Cl, o-Cl, o-NO₂, o-OBn, o-OH, m-F, m,p-diCl, o-pyr,m-pyr and p-pyr. The o-NH₂ analog was obtained by reduction of o-NO₂using H₂/Pt/C in methanol.

EXAMPLE 1104N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](4-methylphenyl)-carboxamide.ES-MS (M+H)+=464, 466 (Cl). EXAMPLE 1105N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](4-methoxyphenyl)-carboxamide.ES-MS (M+H)+=480, 482 (Cl). EXAMPLE 1106N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](4-chlorophenyl)-carboxamide.ES-MS (M+H)+=483.9, 485.9, 487.9 (2Cl). EXAMPLE 1107N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](2-chlorophenyl)-carboxamide.ES-MS (M+H)+=484, 486 (2Cl). EXAMPLE 1108N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](2-nitrophenyl)-carboxamide.ES-MS (M+H)+=495, 497 (Cl). EXAMPLE 1109N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl][2-(phenylmethoxy)-phenyl]carboxamide.ES-MS (M+H)+=556, 558 (Cl). EXAMPLE 1110N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl][2-hydroxyphenyl]-carboxamide.ES-MS (M+H)+=466, 468 (Cl). EXAMPLE 1111N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](3-fluorophenyl)-carboxamide.ES-MS (M+H)+=468, 470 (Cl). EXAMPLE 1112N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](3,4-dichlorophenyl)-carboxamide.ES-MS (M+H)+=517.9, 519.9, 521.9 (3Cl). EXAMPLE 1113N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-2-pyridylcarboxamide.ES-MS (M+H)+=451, 453 (Cl). EXAMPLE 1114N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-3-pyridylcarboxamide.ES-MS (M+H)+=451, 453 (Cl). EXAMPLE 1115N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl]-4-pyridylcarboxamide.ES-MS (M+H)+=451, 453 (Cl). EXAMPLE 1116N-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)-2-methylphenyl](2-aminophenyl)-carboxamide.ES-MS (M+H)+=465, 467 (Cl). EXAMPLE 1117–1120

EXAMPLE 1117

1.0 equivalent ofN-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamidewas treated with 1.0 equivalent of PhCH₂COOH/PyBOP to give X═CH₂,{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{3-methyl-4-[benzylamino]phenyl}carboxamide.ES-MS (M+H)+=464, 466 (Cl).

EXAMPLE 1118

Alternatively (for X═SO₂), the compounds above were prepared accordingto Scheme 4 using the following general synthetic procedure: 1.0equivalents of N-(4-amino-3-methylphenyl){[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide dissolved in DMF (0.5M) was treated with 1.0 equivalent of PhSO₂Cl and 1.2 equivalents ofDIEA to give, after RP-HPLC purification, X═SO₂,{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{3-methyl-4-[(phenylsulfonyl)amino]phenyl}carboxamide.ES-MS (M+H)+=486, 488 (Cl).

EXAMPLE 1119

For X═C═NH: Treatment with methyl benzimidate.HCl (1.4 eq) in DMF gavethe amidine X═C═NH,{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[(iminophenylmethyl)amino]-3-methylphenyl}carboxamide.ES-MS (M+H)+=449, 451 (Cl).

EXAMPLE 1120

For X═NH—C═O: Treatment with phenyl isocyanate (1.07 eq) in DMF to givethe urea X═NH—C═O,{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{3-methyl-4-[(phenylamino)carbonylamino]phenyl}carboxamide.ES-MS (M+H)+=465, 467 (Cl).

EXAMPLE 1121

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(N-phenylcarbamoyl)phenyl]carboxamide A. Synthesis of methyl4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonylamino)benzoate

A solution of 5-chlorothiophene-2-sulfonamide (0.2 g, 1.0 mmol) and DSC(0.307 g, 1.2 mmol) in CH₂Cl₂ (5 μL) and DBU (0.3 mL) was stirred atroom temp overnight. To this mixture was added methyl 4-aminobenzoate(0.15 g, 1.0 mmol). The reaction was then stirred at room temp for 17hr, acidified and HPLC purified to give methyl4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonyl-amino)benzoate (0.23 g,61%). ES-MS (M+H)+=375, 377 (Cl).

B. Synthesis of4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonylamino)benzoic acid

To a suspension of methyl4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonylamino)benzoate (56 mg,0.15 mmol) in methanol (1 mL) and acetonitrile (1 mL) was added 1N LiOH(0.16 mL, 0.16 mmol). The resulting solution was stirred at room tempfor 21 hr, then an additional 0.32 mL 1N LiOH was added and the reactionwas stirred at 40° C. for another 21 hr til complete. Concentration invacuo gave crude4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonylamino)benzoic acid (69mg). ES-MS (M+H)+=361.

C. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-N-phenylcarbamoyl)phenyl]-carboxamide

To a solution of4-({[(5-chloro-2-thienyl)sulfonyl]amino}carbonylamino)benzoic acid (69mg) in DMF (0.7 mL) was added aniline (21 uL, 0.23 mmol), DIEA (3 eq.)then PyBOP (85 mg, 0.16 mmol). The reaction mixture was stirred at roomtemp for 28 hr, acidified and HPLC purified to give{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(N-phenylcarbamoyl)phenyl]carboxamide(29 mg, 45%). ES-MS (M+H)+=436, 438 (Cl).

General Procedure for Synthesizing Isoquinolinone-containing SulfonylUreas

A. General Procedure for synthesis of cinnamic acids

To a solution of malonic acid (10.4 g, 0.1 mol) and the benzaldehyde(0.05 mol) in pyridine (20 mL) was added neat piperidine (0.75 mL, 7.6mmol). The reaction mixture was stirred at 80° C. for 17 hr, chilled,then added to 200 mL cold water. This mixture was acidified with conc.HCl (25 mL) and the white precipitate collected by filtration, washedwith 5×1 0 mL water, and dried to give pure cinnamic acid in typicalyields of >95%.

B-1. General Procedure 1 for Cyclization of Cinnamic Acids toIsoquinolinones

To a chilled suspension of the cinnamic acid (25 mmol) in benzene (40mL) and DMF (5 drops) was added neat thionyl chloride (2.2 mL, 30 mmol).The reaction mixture was stirred at 60° C. for 2 hr, chilled,concentrated in vacuo and dried to give crude acid chloride. To asolution of the acid chloride in 1,2-dichlorobenzene (22 mL) was addedNaN₃ (2.6 g, 40 mmol). After heating at 140° C. for 6 hr, completeconversion to the isocyanate was observed, catalytic I₂ was added andthe reaction was heated to 180° C. for 17 hr. Reaction workups includedeither precipitation of product with hexane or concentration and flashchromatography using EtOAc/CH₂Cl₂ eluent. Yields varied widely dependingon the substituent (5–80%).

B-2. General Procedure 2 for Cyclization of Cinnamic Acids toIsoquinolinones

To a chilled solution of the cinnamic acid (16 mmol) in dry THF (35 mL)and triethylamine (2.9 mL, 20.8 mmol) was added neat ethyl chloroformate(1.85 mL, 19.4 mmol) dropwise over several minutes. The resultingsuspension was stirred cold for 1 hr, then a solution of NaN₃ (1.56 g,24 mmol) in 10 mL water was added. The reaction was stirred at roomtemp. for 1 hr. Reaction workups included either collection of thereaction precipitate, or extraction of product into CH₂Cl₂, giving pureacyl azide in >90% yields. A solution of the acyl azide in1,2-dichlorobenzene (18 mL) was then heated to 140° C. to form theisocyanate, followed by addition of cat. iodine and heating to 180° C.overnight. Workups were the same as in general procedure B-1.

C. General Procedure for Alkylation of Isoquinolinone with1-fluoro-4-nitrobenzene

To a solution of the isoquinolinone (2.5 nmol) in DMF (5 mL) was addedpotassium carbonate (0.7 g, 5 mmol), followed by neatfluoro-4-nitrobenzene (0.3 mL, 2.8 mmol). The reaction mixture wasstirred at 90° C. for 8 hr, poured onto cold water and filtered to givepure product in typical yields of 85–95%.

D. General Reduction Procedure

A suspension of the nitrophenyl intermediate from C (0.75 mmol) andtin(II) chloride dihydrate (0.68 g, 3 mmol) in ethanol (8 mL) wasstirred at 70° C. for 4 hr. The reaction was then chilled, diluted withEtOAc, mixed with Celite, basified with 1M Na₂CO₃ (20 mL) then filtered.The organic layer was washed with water and brine, dried with Na₂SO₄,concentrated in vacuo to give the product aniline in typical yields of85–95%.

E. General Procedure for Coupling the Aniline Intermediate with5-chlorothiophenesulfonamide

To a suspension of 5-chlorothiophene-2-sulfonamide (40 mg, 0.2 mmol) andDSC (61 mg, 0.24 mmol) in dry acetonitrile (1 mL) was added DBU (60 μL,0.4 mmol). The resulting solution was stirred at room temp for 16 hr.The aniline from D (0.2 mmol) was then added as a solid with additionalacetonitrile (1 mL), and the reaction was heated to 70° C. and stirredfor another 17 hr. Acidification and HPLC purification of the crudereaction gave the final target in varying yields (20–70%) depending onthe substituent.

EXAMPLES 371, 372, 374, 376, 379, 380 AND 1122–1128

The compounds above where, for example, R=7-CH₃, 7-Cl, 7-F, 7-CF₃,7-OCH₃, 6-CH_(3,6)-Cl, 6-F, 6-Br, 6-CF₃, 6-OCH₃, and 6,7-diCl weresynthesized from commercially available benzaldehydes or cinnamic acidsusing the general procedure outlined in Scheme 6. The 7,8-diCl analogwas isolated as a by-product during the synthesis of the 6,7-isomer.

EXAMPLE 371{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-methoxy-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=490, 492 (Cl). EXAMPLE 372{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-chloro-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=494, 496, 498 (2Cl). EXAMPLE 374{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-chloro-1-oxo(2-2-hydroisoquinolyl))phenyl]-carboxamide.ES-MS (M+H)+=494, 496, 498 (2Cl). EXAMPLE 376{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-1-oxo(2-2-hydroisoquinolyl))phenyl]-carboxamide.ES-MS (M+H)+=478.0 (Cl). EXAMPLE 379{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-trifluoromethyl-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=528, 530 (Cl). EXAMPLE 380{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methoxy-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=490, 492 (Cl). EXAMPLE 1122{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methyl-1-oxo(2-2-hydroisoquinolyl))phenyl]-carboxamide.ES-MS (M+H)+=473.9, 475.9 (Cl). EXAMPLE 1123{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-methyl-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=474, 476 (Cl). EXAMPLE 1124{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-fluoro-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+H)+=477.9 (Cl). EXAMPLE 1125{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-bromo-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamide.ES-MS (M+1H) +=537.9 (Cl). EXAMPLE 1126{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-trifluoromethyl-1-oxo(2-2-hydroisoquinolyl))-phenyl]-carboxamide.ES-MS (M+H)+=528, 530 (Cl). EXAMPLE 1127N-[4-(6,7-dichloro-1-oxo(2-2-hydroisoquinolyl))phenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=528, 530, 532 (3Cl). EXAMPLE 1128N-[4-(7,8-dichloro-1-oxo(2-2-hydroisoquinolyl))phenyl]{[(5-chloro(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=528, 530, 532 (3Cl). EXAMPLE 1129

Preparation of{[(5-chloro(2-thienyl))sulfonyl]ammio}-N-[4-(7-chloro-6-methoxy-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamide

A. Synthesis of 4-chloro-3-methoxytoluene

To a solution of 2-chloro-5-methylphenol (10.7 g, 75 mmol) in DMF (40mL) was added potassium carbonate (26 g, 188 mmol) followed by neatmethyl iodide (4.9 mL, 79 mmol). The reaction mixture was stirred atroom temp for 6 hr, extracted with EtOAc, washed with water and brine,dried and concentrated in vacuo to give 4-chloro-3-methoxytoluene (10.8g, 92%). ¹H-NMR (DMSO-d₆): δ 2.26 (s, 3H), 3.79 (s, 3H), 6.71–6.73 (dd,1H), 6.94 (s, 1H), 7.22-7.24 (d, 1H).

B. Synthesis of 4-chloro-3-methoxybenzoic acid

To the crude toluene (7.8 g, 50 mmol) was added a solution KMnO₄ (19.8g, 125 mmol) in water (300 mL). The reaction mixture was stirredvigorously at reflux for 17 hr and filtered warm through Celite, washingthe cake with 200 mL hot water. The clear filtrate was washed with ethylether (2×150 mL), acidified with conc. HCl (9 mL) and filtered to givepure white solid 4-chloro-3-methoxybenzoic acid (5.36 g, 58%). ES-MS(M+H)+=187.

C. Synthesis of (4-chloro-3-methoxyphenyl)methan-1-ol

To a solution of 4-chloro-3-methoxybenzoic acid (4.88 g, 26.2 mmol) inTHF (50 mL) was added borane-THF complex (52 mL 1M solution in THF, 52mmol) via addition funnel over 10 min. The reaction mixture was refluxedfor 2 hr, cooled, extracted with EtOAc, washed with water, 5% Na₂CO₃ andbrine, dried and concentrated in vacuo to give(4-chloro-3-methoxyphenyl)methan-1-ol (4.5 g, 99%). ES-MS(M+H—H₂O)+=155, 157 (Cl).

D. Synthesis of 4-chloro-3-methoxybenzaldehyde

To a solution of (4-chloro-3-methoxyphenyl)methan-1-ol (5.08 g, 29.4mmol) in benzene (120 mL) was added MnO₂ (5.65 g, 65 mmol). The reactionmixture was refluxed for 17 hr, chilled, and filtered through Celite,washing the cake with CH₂Cl₂ (300 mL). The filtrate was concentrated invacuo to give 4-chloro-3-methoxybenzaldehyde (4.5 g, 89%).

E. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-chloro-6-methoxy-1-oxo(2-2-hydroisoguinolyl))phenyl]carboxamide

{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-chloro-6-methoxy-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamidewas synthesized from the benzaldehyde using the general procedureoutlined in Scheme 6, steps A–E. ES-MS (M+H)+=489, 491 (Cl).

EXAMPLE 1130

Preparation ofN-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]-1-oxo-6-2-hydroisoquinolyl}acetamide A. Synthesis ofN-(3-formylphenyl)acetamide

To a chilled suspension of 3-aminobenzyl alcohol (9.24 g, 75 mmol) inTHF (50 mL) was added neat acetic anhydride (8.1 mL, 86 mmol). Thereaction mixture was stirred cold for 1 hr, diluted with EtOAc, washedwith aq. NaOH and brine, and concentrated in vacuo to giveN-[3-(hydroxymethyl)phenyl]acetamide (10.5 g, 85%).

A mixture of N-[3-(hydroxymethyl)phenyl]acetamide (10 g, 60.6 mmol) andMnO₂ (7.8 g, 90 mmol) in toluene (250 mL) was refluxed for 29 hr, withaddition of more Mn02 (0.7 g, 9 mmol) at 24 hr. The reaction was cooled,filtered through Celite and concentrated in vacuo to giveN-(3-formylphenyl)acetamide (9.2 g, 75%). ES-MS (M+H)+=164.

B. Synthesis ofN-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]-1-oxo-6-2-hydroisoguinolyl}acetamide

N-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]-1-oxo-6-2-hydroisoquinolyl}acetamidewas synthesized from N-(3-formylphenyl)acetamide using the generalprocedure outlined in Scheme 6, steps A–E. ES-MS (M+H)+=517, 519 (Cl).

EXAMPLE 1131

Preparation ofN-[4-(6-amino-1-oxo(2-2-hydroisoquinolyl))phenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}-carboxamide:This compound was synthesized by treatingN-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonyl-amino)phenyl]-1-oxo-6-2-hydroisoquinolyl}acetamidewith 30 equiv. NaOMe in MeOH and refluxing overnight. It was alsosynthesized by treatment with neat hydrazine hydrate at 70° C. ES-MS(M+H)+=475, 477 (Cl).

EXAMPLE 373

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylamino)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamideA. Synthesis ofN-methyl-N-[2-(4-nitrophenyl)-1-oxo(6-2-hydroisoguinolyl)]acetamide

To a solution of crudeN-[2-(4-nitrophenyl)-1-oxo-6-2-hydroisoquinolyl]acetamide (0.26 g, 0.8mmol) in DMF (2 mL) was added cesium carbonate (0.645 g, 2 mmol)followed by neat methyl iodide (75 μL, 1.2 mmol). The reaction mixturewas stirred at room temp for 17 hr, precipitated with addition of waterand filtered to giveN-methyl-N-[2-(4-nitrophenyl)-1-oxo(6-2-hydroisoquinolyl)]acetamide (75mg, 25%). ES-MS (M+H)+=338.

B. Synthesis ofN-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]-1-oxo(6-2-hydroisoquinolyl)}-N-methylacetamide

The reduction and coupling steps were performed using the proceduresoutlined in Scheme 6, steps D and E. ES-MS (M+H)+=531, 533 (Cl).

C. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylamino)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamide

To a solution ofN-{2-[4-({[(5-chloro(2-thienyl))sulfonyl]amino}carbonylamino)phenyl]-1-oxo(6-2-hydroisoquinolyl)}-N-methylacetamide(35 mg, 0.072 mmol) in methanol (1.2 mL) was added 0.5 M NaOMe (0.44 mL,0.22 mmol) in methanol. The reaction mixture was stirred at 60° C.overnight, acidified and HPLC purified to give{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylamino)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamide(22 mg, 63%). ES-MS (M+H)+=489, 491 (Cl).

EXAMPLES 383 AND 1132–1135

The compounds above where, for example, R=Et, n-Pr, CH₂-c-r, CH₂CH₂F andbenzyl were synthesized using the procedure outlined in Example 373 forR=Me, varying the alkylating agent in step A.

EXAMPLE 383{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(ethylamino)-1-oxo(2-2-hydroisoquinolyl)]-phenyl}carboxamide.ES-MS (M+H)+=503, 505 (Cl). EXAMPLE 1132{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[1-oxo-6-(propylamino)(2-2-hydroisoquinolyl)]-phenyl}carboxamide.ES-MS (M+H)+=517, 519 (Cl). EXAMPLE 1133{[(5-chloro(2-thienyl))sulfonyl]amino}-N-(4-{6-[(cyclopropylmethyl)amino]-1-oxo(2-2-hydroisoquinolyl)}phenyl)carboxamide.ES-MS (M+H)+=529, 531 (Cl). EXAMPLE 1134{[(5-chloro(2-thienyl))sulfonyl]amino}-N-(4-{6-[(2-fluoroethyl)amino]-1-oxo(2-2-hydroisoquinolyl)}phenyl)carboxamide.ES-MS (M+H)+=521, 523 (Cl). EXAMPLE 1135{[(5-chloro(2-thienyl))sulfonyl]amino}-N-(4-{1-oxo-6-[benzylamino](2-2-hydroisoquinolyl)}-phenyl)carboxamide.ES-MS (M+H)+=565, 567 (Cl). EXAMPLE 1136

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[1-oxo-6-(phenylamino)(2-2-hydroisoquinolyl)]phenylcarboxamide A.2-(4-nitrophenyl)-6-(phenylamino)-2-hydroisoquinolin-1-one

To a dry RBF under argon was added6-bromo-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one (66 mg, 0.191 mmol)(prepared as outlined in Example 1125), cesium carbonate (106 mg, 0.325mmol), tris(dibenzylideneacetone) dipalladium(0) (3.5 mg, 0.076 mmol),9,9-dimethyl-4,5-bis(diphenylphosphino) xanthene (12 mg, 0.0207 mmol)and neat aniline (0.026 mL, 0.285 mmol). To this flask was added drydioxane (0.5 mL) and dry toluene (0.5 mL). The reaction was stirred at80° C. for 5 hr, concentrated and chromatographed on silica gel to givepure 2-(4-nitrophenyl)-6-(phenylamino)-2-hydroisoquinolin-1-one (55 mg,81%). ES-MS (M+H)+=358.

B.{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4[-1-oxo-6-(phenylamino)(2-2-hydroisoquinolyl)]phenyl}carboxamide

Preparation of the final target was accomplished using the generalprocedure outlined in Scheme 6, steps D–E, to give the above namedsulfonyl urea. ES-MS (M+H)+=551, 553 (Cl).

EXAMPLE 1137

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-(4-{6-[(2-methoxyethyl)amino]-1-oxo(2-2-hydroisoquinolyl)}phenyl)carboxamidewas accomplished using a similar Buchwald procedure as shown in Example1136. ES-MS (M+H)+=533.0 (Cl).

EXAMPLE 1138

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-hydrazino-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamide

A 5 mg (0.01 mmol) sample of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-fluoro-1-oxo(2-2-hydroisoquinolyl))-phenyl]carboxamidewas dissolved in 50 μL of neat anhydrous hydrazine and stirred for 18 h.The solution was diluted with 250 uL of water and lyopholized to give3.8 mg (74%) of the desired material. ES-MS (M+H)+=490.0 (Cl).

EXAMPLE 1139

Preparation ofN-{4-[6-(dimethylamino)-1-oxo(2-2-hydroisoquinolyl)]phenyl}{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide:To a suspension of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylamino)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamide(17 mg, 0.035 mmol) (prepared in Example 373) in glacial acetic acid(0.7 mL) was added formaldehyde (12 μL, 0.15 mmol) followed by sodiumtriacetoxyborohydride (14 mg, 0.067 mmol). The reaction mixture wasstirred at 45° C. for 2 hr. HPLC purification yielded the final product(7 mg, 40%). ES-MS (M+H)+=503, 505 (Cl).

EXAMPLE 1140

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-hydroxy-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamideA. Synthesis of 6-hydroxy-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one

To a solution of 6-methoxy-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one(100 mg, 0.338 mmol) (prepared by the general procedure outline inScheme 6) in CH₂Cl₂ (3 mL) was added a 1M BBr₃ solution in CH₂Cl₂ (1.35mL, 1.35 mmol). The solution was refluxed for 18 h, the solvent wasremoved in vacuo, the residue was triturated with water, and theresulting greenish solid was collected and dried to give6-hydroxy-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one (89 mg, 93%).

B. Synthesis of 2-(4-aminophenyl)-6-hydroxy-2-hydroisoquinolin-1-one

6-hydroxy-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one was reducedaccording to the general procedure in Scheme 6 to give the correspondinganiline in 27% yield. ES-MS (M+H)+=252.9.

C. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(6-hydroxy-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamide

2-(4-aminophenyl)-6-hydroxy-2-hydroisoquinolin-1-one was coupledaccording to the general procedure outlined in Scheme 6 to give theabove named sulfonyl urea. ES-MS (M+H)+=476 (Cl).

EXAMPLE 1141

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylethoxy)-1-oxo(2-hydroisoquinolyl)]phenyl}carboxamideA. Synthesis of6-(methylethoxy)-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one

To a solution of 6-hydroxy-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one(50 mg, 0.177 mmol) in DMF (0.38 mL) was added 2-bromopropane (0.03 mL)and cesium carbonate (86 mg, 0.267 mmol). After heating at 60° C. for 18h, water was added and the solution stirred and cooled to 0° C. Theprecipitate was collected and dried to give6-(methylethoxy)-2-(4-nitrophenyl)-2-hydroisoquinolin-1-one (34 mg,59%).

B. Synthesis of2-(4-aminophenyl)-6-(methylethoxy)-2-hydroisoquinolin-1-one

This material was reduced according to the general procedure in Scheme 6to give the corresponding aniline in 91% yield. ES-MS (M+H)+=295.

C. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(methylethoxy)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamide

2-(4-aminophenyl)-6-(methylethoxy)-2-hydroisoquinolin-1-one was coupledaccording to the general procedure outlined in Scheme 6 to give theabove named sulfonyl urea. ES-MS (M+H)+=518 (Cl); ES-MS (M−H)+=516.

EXAMPLE 1142

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-{4-[6-(2-methoxyethoxy)-1-oxo(2-2-hydroisoquinolyl)]phenyl}carboxamidewas accomplished according to the procedure of Example 1141 to give thedesired sulfonyl urea. ES-MS (M+H)+=534.1 (Cl).

EXAMPLE 1143

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methylthio-1-oxo(2-2-hydroisoquinolyl))phenyl]carboxamidewas accomplished according to the procedure of Example 1145, using{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-1-oxo(2-2-hydroisoquinolyl))phenyl]-carboxamideas starting material, to give the desire sulfonyl urea. ES-MS (M+H)+=506(Cl).

EXAMPLE 1144

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamideA. Synthesis of ethyl 2-amino-4-fluorobenzoate

To a chilled solution of 2-amino-4-fluorobenzoic acid (1.57 g, 10.1mmol) in absolute ethanol (20 mL) was added neat thionyl chloride (4.4mL, 60 mmol). The reaction mixture was refluxed for 4 days total, withaddition of more SOCl₂ (8 mL, 110 mmol), then concentrated, diluted withEtOAc, washed with 2N NaOH, dried and concentrated in vacuo to giveethyl 2-amino-4-fluorobenzoate (1.73 g, 94%).

B. Synthesis of 7-fluoro-3-hydroquinazolin-4-one

To a suspension of ethyl 2-amino-4-fluorobenzoate (1.73 g, 9.45 mmol) informamide (8 mL) was added ammonium formate (0.9 g, 14 mmol). Thereaction mixture was stirred at 140° C. for 24 hr, with additionalammonium formate (0.92 g, 15 mmol) at 6 hr. The reaction was dilute withEtOAc, washed with water, back-extracted with EtOAc, dried andconcentrated in vacuo to give 7-fluoro-3-hydroquinazolin-4-one (2.82 g)which contains some formamide. ES-MS (M+H)+=165.

C. Synthesis of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-4-oxo(3-hydroquinazolin-3-yl]phenyl}carboxamide

{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamidewas synthesized from the quinazolinone using the procedures foralkylation, reduction and coupling outlined in Scheme 6, steps C, D andE. The alkylation product was chromatographed on silica gel to removeformamide carried over from the previous step to give pure intermediatein 42% yield. The reduction step was performed in EtOAc instead of EtOH.The coupling proceeded in 50% yield. ES-MS (M+H)+=479, 481 (Cl).

EXAMPLE 507

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methoxy-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamide:To a solution of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-4-oxo(3-hydro-quinazolin-3-yl))phenyl]carboxamide(20 mg, 0.042 mmol) in methanol (0.75 mL) and DMF (0.3 mL) was added 0.5M NaOMe in MeOH (0.42 mL, 0.21 mmol). The reaction mixture was stirredat 70° C. for 24 hr, acidified and HPLC purified to give pure{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methoxy-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamide(7 mg, 33%). ES-MS (M+H)+=491, 493 (Cl).

EXAMPLE 1145

Preparation of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methylthio-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamide:To a solution of{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-fluoro-4-oxo(3-hydro-quinazolin-3-yl))phenyl]carboxamide(20 mg, 0.042 mmol) in DMF (0.21 mL) was added NaSMe (7 mg, 0.1 mmol).The reaction mixture was stirred at room temp for 3 hr, acidified andHPLC purified to give pure{[(5-chloro(2-thienyl))sulfonyl]amino}-N-[4-(7-methylthio-4-oxo(3-hydroquinazolin-3-yl))phenyl]carboxamide(17 mg, 80%). ES-MS (M+H)+=507, 509 (Cl).

EXAMPLE 1146

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))-3-bromophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamideA. Synthesis of3-(2-bromo-4-nitrophenyl)-1,3-dihydroquinazoline-2,4-dione

To a solution of methyl 2-isocyanatobenzoate (0.266 g, 1.5 mmol) and2-bromo-4-nitroaniline (0.325 g, 1.5 mmol) in DMF (2 mL) was added DIEA(0.79 mL). The reaction mixture was stirred at room temp for 24 hr, withaddition of DBU (0.22 mL) at 17 hr. The reaction mixture was extractedwith EtOAc, washed with 1 N HCl and brine, dried and concentrated togive crude product, which was chromatographed on silica gel with 10%EtOAc/CH₂Cl₂ to give3-(2-bromo-4-nitrophenyl)-1,3-dihydroquinazoline-2,4-dione (0.24 g,44%). ES-MS (M+H)+=362, 364 (Br).

B. Synthesis of3-(4-amino-2-bromophenyl)-1,3-dihydroquinazoline-2,4-dione

A suspension of3-(2-bromo-4-nitrophenyl)-1,3-dihydroquinazoline-2,4-dione (0.18 g, 0.5mmol) and tin(II) chloride dihydrate (0.45 g, 2.0 mmol) in ethyl acetate(5 mL) was heated at 70° C. for 4 hr. The reaction mixture was thencooled, mixed with Celite, made basic with 4N NaOH (3 mL), filteredthrough Celite, and concentrated in vacuo to give the desired compound(0.155 g, 94%). ES-MS (M+H)+=332, 334 (Br).

C. General Procedure for Coupling Anilines with Aryl Sulfonamides toForm Sulfonyl Ureas

A solution of the aryl sulfonamide (0.15 mmol) and DSC (0.18 mmol) inCH₂Cl₂ (1 mL) and DBU (45 μL, 0.3 mmol) was stirred at room temp for 16hr. To this mixture was added the aniline intermediate (0.15 mmol) andCH₃CN (1 mL) and DBU (23 μL, 0.15 mmol) (if aniline is TFA salt). Thereaction was heated at 60° C. for 17 hr, acidified and HPLC purified togive sulfonyl urea product in typical yields between 25–70%.

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))-3-bromophenyl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamidewas achieved in 25% yield. ES-MS (M+H)+=556.9. 558.9 (Br, Cl).

EXAMPLE 302

Preparation ofN-[6-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(3-pyridyl)]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

This compound was prepared by first following step A in Example 1146,reacting 2-amino-5-nitropyridine with methyl 2-isocyanatobenzoate. Thenitro group was reduced under 1 atm H₂, 10% Pd/C, 1 eq. HCl, MeOHconditions for 6 hr. After filtration and concentration, the aniline wascoupled with 5-chlorothiophene-2-sulfonamide using the conditionsoutlined in step C in Example 1146 to giveN-[6-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(3-pyridyl)]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide(47 mg, 33%). ES-MS (M+H)+=478, 480 (Cl).

EXAMPLE 1147

Preparation of 3-(4-aminophenyl)-1,3-dihydroquinazoline-2,4-dionetrifluoroacetate salt A. Synthesis of(tert-butoxy)-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide

To a solution of methyl 2-isocyanatobenzoate (0.97 g, 5.5 mmol) and Boc1,4-phenylenediamine (1.04 g, 5 mmol) in THF (15 mL) was added DIEA(0.87 mL, 5 mmol) and DBU (0.75 mL, 5 mmol). The reaction mixturestirred at room temp for 5 hr, the off-white solid filtered and washedwith ethyl ether to give desired compound (1.49 g, 85%). ES-MS(M+Na)+=376.1, (M-tBu+H)+=298.0.

B. Synthesis of 3-(4-aminophenyl)-1,3-dihydroquinazoline-2,4-dionetrifluoroacetate salt

To a chilled suspension of(tert-butoxy)-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide(0.35 g, 1 mmol) in CH₂Cl₂ (2 mL) was added neat TFA (2 mL). Theresulting solution was stirred cold for 1 hr, concentrated in vacuo,azeotroped with heptane and dried to give desired compound (0.376 g,99%) as the mono TFA salt. ES-MS (M+H)+=254.

Synthesis of Various Aryl Sulfonylurea Analogs

The sulfonyl urea targets above were prepared using the procedureoutlined in Example 1146 Step C, reacting the aniline from Example 1146with the following 13 commercially available sulfonamides:5-nitrothiophene-2-sulfonamide; thiophene-2-sulfonamide;5-chloro-3-methylbenzothiophene-2-sulfonamide;3,5-dimethylisoxazole-4-sulfonamide;N-(3-methyl-5-sulfamoyl)-3H(1,3,4)thiadiazol-2-ylidene)acetamide;2,4-dimethyl-1,3-thiazole-5-sulfonamide;3-bromo-5-chlorothiophene-2-sulfonamide; azetazolamide;5-isoxazol-3-ylthiophene-2-sulfonamide; 2-chlorobenzenesulfonamide;3-chlorobenzenesulfonamide; 4-methoxybenzenesulfonamide;4-(trifluoromethyl)benzenesulfonamide.

EXAMPLE 1148N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-nitro(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=488. EXAMPLE 1149

N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(2-thienylsulfonyl)amino]carboxamide.ES-MS (M+H)+=443.0.

EXAMPLE 1150N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-chloro-3-methyl-benzo-[b]thiophen-2-yl)sulfonyl]amino}carboxamide.ES-MS (M+H)+=541, 543 (Cl). EXAMPLE 1151{[(3,5-dimethylisoxazol-4-yl)sulfonyl]amino}-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide.ES-MS (M+H)+=456. EXAMPLE 1152({[2-(1-aza-2-oxopropylidene)-3-methyl(1,3,4-thiadiazolin-5-yl)]sulfonyl}amino)-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide.ES-MS (M+H)+=516. EXAMPLE 1153{[(2,4-dimethyl(1,3-thiazol-5-yl))sulfonyl]amino}-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide.ES-MS (M+H)+=472. EXAMPLE 1154N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(3-bromo-5-chloro(2-thienyl))-sulfonyl]amino}carboxamide.ES-MS (M+H)+=554.8, 556.9, 558.8 (BrCl). EXAMPLE 1155N-{5-[({N-[4-(2,4-dioxo-1,3-dihydroquinazolin-3-yl)phenyl]carbamoyl}amino)sulfonyl]-1,3,4-thiadiazol-2-yl}acetamide.ES-MS (M+H)+=502. EXAMPLE 1156N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-isoxazol-3-yl(2-thienyl))-sulfonyl]amino}carboxamide.ES-MS (M+H)+=510. EXAMPLE 1157N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(2-chlorophenyl)sulfonyl]amino}-carboxamide. ES-MS (M+H)+=471, 473 (Cl). EXAMPLE 1158N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(3-chlorophenyl)sulfonyl]amino}carboxamide.ES-MS (M+H)+=471, 473 (Cl) EXAMPLE 1159N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(4-methoxyphenyl)sulfonyl]amino}-carboxamide.ES-MS (M+H)+=467. EXAMPLE 1160N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]({[4-(trifluoromethyl)phenyl]sulfonyl}amino)carboxamide.ES-MS (M+H)+=505

An additional 6 sulfonamides were prepared from the followingcommercially available sulfonyl chlorides, and were subsequently coupledwith the aniline using the procedure in Example 1146 step C:2-acetamido-4-methyl-5-thiazolesulfonyl chloride;4-fluorobenzenesulfonyl chloride; 5-(pyrid-2-yl)thiophene-2-sulfonylchloride; 3,4-dichlorobenzenesulfonyl chloride;2-(trifluoromethyl)benzenesulfonyl chloride;3-(trifluoromethyl)benzenesulfonyl chloride.

EXAMPLE 1161N-{5-[({N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carbamoyl}amino)sulfonyl]-4-methyl-1,3-thiazol-2-yl}acetamide.ES-MS (M+H)+=515. EXAMPLE 1162N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(4-fluorophenyl)sulfonyl]amino}-carboxamide. ES-MS (M+H)+=455.1. EXAMPLE 1163N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-(2-pyridyl)(2-thienyl))sulfonyl]-amino}carboxamide.ES-MS (M+H)+=520. EXAMPLE 1164{[(3,4-dichlorophenyl)sulfonyl]amino}-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide.ES-MS (M+H)+=505, 507 (2Cl). EXAMPLE 1165N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]({[2-(trifluoromethyl)phenyl]sulfonyl}-amino)carboxamide.ES-MS (M+H)+=505. EXAMPLE 1166N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]({[3-(trifluoromethyl)phenyl]-sulfonyl}amino)carboxamide.ES-MS (M+H)+=505. EXAMPLE 1167

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(1,3-thiazol-2-ylsulfonyl)amino]carboxamideA. Synthesis of (tert-butyl(1,3-thiazol-2-ylsulfonyl)amine

To a suspension of 2-mercaptothiazole (0.16 g, 1.37 mmol) in CH₂Cl₂ (14mL) was added water (7 mL) followed by N-chlorosuccinimide (0.75 g, 5.6mmol). The reaction mixture was stirred vigorously for 1.5 hr, dilutedwith CH₂Cl₂, washed with sat. NaHCO₃, water and brine, dried andconcentrated in vacuo to give the crude sulfonyl chloride (0.25 g). Asolution of the sulfonyl chloride and t-butylamine (0.75 mL, 7.1 mmol)in THF (2.5 mL) was stirred at room temp for 3 hr. The reaction wasdiluted with EtOAc, washed with 1N HCl, water and brine, dried andconcentrated in vacuo to give desired compound (0.16 g, 53%).

B. Synthesis of 1,3-thiazole-2-sulfonamide

To a solution of (tert-butyl)(1,3-thiazol-2-ylsulfonyl)amine (0.22 g,1.0 mmol) in 1,2-dichloroethane (10 mL) was added methanesulfonic acid(0.26 mL, 4 mmol). The reaction mixture was heated at 80° C. for 9 hr,concentrated in vacuo and chromatographed to give pure sulfonamide (0.14g, 88%). ES-MS (M+H)+=164.9.

C. Synthesis ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(1,3-thiazol-2-ylsulfonyl)amino]carboxamide

The sulfonyl urea was prepared by coupling the aniline from Example 1147with 1,3-thiazole-2-sulfonamide using the procedure outlined in Example1146 step C. ES-MS (M+H)+=444.0.

EXAMPLE 1168

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(5-chloro-1,3-thiazol-2-ylsulfonyl)amino]carboxamideA. Synthesis of (tert-butyl)[(5-chloro(1,3-thiazol-2-yl))sulfonyl]amine

To a solution of (tert-butyl)(1,3-thiazol-2-ylsulfonyl)amine (0.15 g,0.7 mmol) in ethyl ether (3 mL) at −78° C. was added a 1.6M solution ofn-butyllithium (0.875 mL, 1.4 mmol) in hexanes via syringe under argon.The reaction mixture was stirred at −7,8° C. for 1 hr, then neatbenzenesulfonyl chloride (90 μL, 0.7 mmol) was added. The resultingsuspension was stirred at room temp for 2 hr. The reaction was dilutedwith EtOAc, washed with water and brine, dried, concentrated in vacuoand chromatographed (15% EtOAc/hexane) to give pure desired compound (58mg, 33%). ES-MS (M+Na)+=277, 279 (Cl), (M-tBu+H)+=199, 201 (Cl).

B. Synthesis of 5-chloro-1,3-thiazole-2-sulfonamide

To a solution of (tert-butyl)[(5-chloro(1,3-thiazol-2-yl))sulfonyl]amine(56 mg, 0.22 mmol) in 1,2-dichloroethane (2 mL) was addedmethanesulfonic acid (50 μL, 0.77 mol). The reaction mixture was heatedat 80° C. for 3 hr, concentrated in vacuo and chromatographed (30%EtOAc/hexane) to give pure sulfonamide (42 mg, 96%). ES-MS (M+H)+=199(Cl).

C. Synthesis ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(5-chloro-1,3-thiazol-2-ylsulfonyl)amino]carboxamide

The sulfonyl urea was prepared by coupling the aniline from Example 1147with 5-chloro-1,3-thiazole-2-sulfonamide using the procedure outlined inExample 1146 step C. ES-MS (M+H)+=478, 480 (Cl).

EXAMPLE 1169

Preparation of[(benzo[b]thiophen-2-ylsulfonyl)amino]-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamideA. Synthesis of benzo[b]thiophene-2-sulfonamide

To a solution of benzothiophene (1.63 g, 12.1 mmol) in THF (8 mL) at 0°C. was added a 1.6M solution of n-butyllithium (8.5 mL, 13.6 mmol) inhexanes slowly over 10 min via syringe. The reaction was stirred coldfor 10 min. THF (8 μL) was added and the entire reaction was transferredvia cannula to a vessel containing sulfuryl chloride (2 mL, 25 mmol) inhexane (8 mL) at 0° C. The resulting yellow suspension was stirred at 0°C. for 1 hr and eventually became a clear yellow solution. This solutionwas concentrated to about 10 mL volume, diluted with acetone (12 mL) andadded to a solution of ammonium hydroxide (8 mL) in acetone (25 mL). Thereaction mixture was stirred at room temp for 2 hr, added to 200 mL ofwater on ice bath, acidified with conc. HCl (6 mL). A precipitate wasfiltered to obtain light yellow solid (1.78 g). This crude product wasdissolved in 0.5 N KOH (100 mL) and washed with ethyl ether (50 mL).Upon acidification with conc. HCl (6 mL), the product was extracted intoEtOAc (2×60 mL), washed with water and brine, dried and concentrated invacuo to give pure sulfonamide (0.99 g, 39%).

B. Synthesis of[(benzo[b]thiophen-2-ylsulfonyl)amino]-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide

The sulfonyl urea was prepared by coupling the aniline from Example 1147with benzo[b]thiophene-2-sulfonamide using the procedure outlined inExample 1146 step C. ES-MS (M+H)+=493.

EXAMPLE 1170

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-methoxy(2-thienyl))sulfonyl]amino}carboxamideA. Synthesis of 5-methoxythiophene-7-2-sulfonamide

To a solution of 2-methoxythiophene (1 mL, 10 mmol) in dry THF (36 mL)at 78° C. was added a 1.6 M solution of n-butyllithium (8 mL, 12.8 mmol)in hexanes over 10 min via syringe. The reaction was stirred at −78° C.for 2 hr. SO₂ (gas) was bubbled into the reaction mixture for about 10min, then the reaction was allowed to come to room temp and stirred for1 hr. A solution of sodium acetate (6.56 g, 80 mmol) andhydroxylamine-O-sulfonic acid (3.14 g, 27.8 mmol) in water (40 mL) wasthen added, and the reaction was stirred vigorously for 2 hr. Thereaction was basified with 4N NaOH (15 mL), washed with ethyl ether,acidified with 6N HCl (15 mL), extracted with CH₂Cl₂, washed with waterand brine, dried and concentrated in vacuo to give pure sulfonamide(1.01 g, 53%). ES-MS (M+H)+=194.

B. Synthesis ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-methoxy(2-thienyl))sulfonyl]aminocarboxamide

The sulfonyl urea was prepared by coupling the aniline from Example 1147with 5-methoxythiophene-2-sulfonamide using the procedure outlined inExample 1146 step C. ES-MS (M+H)+=473.

EXAMPLE 1171

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(1,1-dioxobenzo[d]thiol-2-yl)sulfonyl]amino}carboxamideA. Synthesis ofN-[4-(1,3-dioxobenzo[c]azolidin-2-yl)naphthyl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

To a solution of the sulfonamide from Example 7 (0.213 g, 1 mmol) inCH₂Cl₂ (4 mL) was added m-chloroperbenzoic acid (0.49 g, 2.2 mmol). Thereaction mixture was refluxed for 20 hr, diluted with EtOAc, washed with5% NaHCO₃, 1N HCl and brine, dried and concentrated in vacuo to givesulfonamide (0.17 g, 71%). ES-MS (M+H)+=246.

B. Synthesis ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]}[(1,1-dioxobenzo[d]thiol-2-yl)sulfonyl]amino}carboxamide

The sulfonyl urea was prepared by coupling the aniline from Example 1147withN-[4-(1,3-dioxobenzo[c]azolidin-2-yl)naphthyl]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamideusing the procedure outlined in Example 1146 step C. ES-MS (M+H)+=525.

EXAMPLE 1172

Preparation of{[(5-amino(2-thienyl))sulfonyl]amino}-N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]carboxamide

To a solution ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-nitro(2-thienyl))sulfonyl]amino}carboxamide(from Example 1148) (20 mg, 0.041 mmol) in methanol (1.5 mL) andtriethylamine (11 μL, 0.08 mmol) was added 10% Pd/C (5 mg, 0.005 mmol)under argon. The reaction mixture was hydrogenated under 1 atm H₂ for 3hr, filtered, concentrated and HPLC purified to give the aniline (6 mg,33%). ES-MS (M+H)+=458.

EXAMPLE 1173

Preparation ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(3-methylbenzo[b]thiophen-2-yl)sulfonyl]amino}carboxamide

To a solution of the sulfonylurea from Example 1150 (52 mg, 0.046 mmol)in methanol (1.5 mL) and triethylamine (12 μL) was added 10% Pd/C (50mg) and PtO₂ (7 mg). The reaction mixture was hydrogenated under 250 psiH₂ for 4 days, filtered, concentrated and HPLC purified to give thedehalogenated product (2 mg, 10%). ES-MS (M+H)+=507.

EXAMPLES 1174–1176

The sulfonyl urea targets above were prepared by reaction of the aniline3-(4-aminophenyl)-1,3-dihydroquinazoline-2,4-dione trifluoroacetate salt(Example 1147) with 3 commercially available substitutedphenylsulfonylisocyanates (1.5 eq.) (R═H, Cl, CH₃) in DMF. Products weretypically isolated by precipitation from DMF reaction mixture with waterand filtration.

EXAMPLE 1174N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl][(phenylsulfonyl)amino]carboxamide.ES-MS (M+H)+=437.0. EXAMPLE 1175N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(4-chlorophenyl)sulfonyl]amino}carboxamide.ES-MS (M+H)+=471, 473 (Cl). EXAMPLE 1176N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(4-methylphenyl)sulfonyl]amino}carboxamide.ES-MS (M+H)+=451. EXAMPLE 1177

Preparation ofN-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamideA. Synthesis of (tert-butoxy)-N-(5-nitro(2-pyridyl))carboxamide

To a solution of 2-amino-5-nitropyridine (0.555 g, 4 mmol) in THF (10mL) was added 1M NaHMDS in THF (8 mL, 8 mmol). The resulting dark redsuspension was stirred for 15 min, followed by addition of a solution ofBoc anhydride (0.87 mL, 3.8 mmol) in THF (5 mL). The reaction mixturewas stirred at room temp for 21 hr, dilute with EtOAc, washed with 1NHCl and brine, dried and concentrated in vacuo to give desired compound(0.63 g, 70%). ES-MS (M+H)+=240, (M-tBu+H)+=184.

B. Synthesis of N-(5-amino(2-pyridyl))(tert-butoxy)carboxamide

To a suspension of (tert-butoxy)-N-(5-nitro(2-pyridyl))carboxamide (0.27g, 1.13 mmol) in methanol (2 mL), ethyl acetate (4 mL) and TEA (0.16 mL)was added 10% Pd/C (60 mg, 0.056 mmol) under argon. The reaction mixturewas hydrogenated under 1 atm H₂ for 20 hr, filtered through Celite andconcentrated in vacuo to give desired compound (0.226 g, 97%).

C. Synthesis of[(tert-butyl)amino]-N-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)]carboxamide

The above named compound was prepared using the procedure outlined inExample 1146 step A by reaction ofN-(5-amino(2-pyridyl))(tert-butoxy)carboxamide with methyl2-isocyanatobenzoate. ES-MS (M+H)+=355.

D. Synthesis ofN-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)]{[(5-chloro(2-thienyl))sulfonyl]amino}carboxamide

To a solution of 5-chlorothiophene-2-sulfonamide (20 mg, 0.1 mmol) and[(tert-butyl)amino]-N-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)]carboxamide(35 mg, 0.1 mmol) in DMF (1 mL) was added DBU (30 μL). The reactionmixture was heated at 90° C. for 3 days, acidified and HPLC purified togive the sulfonyl urea (7 mg, 16%). ES-MS (M+H)+=478, 480 (Cl).

EXAMPLE 1178

Preparation ofN-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)][(phenylsulfonyl)amino]carboxamide

This compound was prepared by TFA deprotection of[(tert-butyl)amino]-N-[5-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))(2-pyridyl)]carboxamidefrom Example 1177, step C, followed by reaction of the aminopyridinewith phenylsulfonylisocyanate (1.5 eq) in DMF, precipitation from 0.1%TFA and filtration of product to give the sulfonyl urea desired compound(40% yield). ES-MS (M+H)+=438.

EXAMPLE 1179

N-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-chloro-3-fluoro(2-thienyl))sulfonyl]amino}carboxamide

A. Synthesis of (tert-butyl)[(5-chloro(2-thienyl))sulfonyl]amine

A solution of 5.5 g (27.5 mmol) of 5-chlorothiophenesulfonyl chloride indry THF at 0° C. was treated with a solution of 5.7 mL (75.5 mmol) oft-butylamine. After warming to 23° C., the reaction mixture was dilutedwith 125 mL of diethyl ether, filtered, and washed with 1 N HCl, brine,and dried (MgSO₄). Concentration in vacuo affords 6.4 g (98%) of thenamed compound as an oil.

B. Synthesis of(tert-butyl)[(5-chloro-3-fluoro(2-thienyl))sulfonyl]amine

A THF solution (1.5 mL) of 128 mg (0.50 mmol) of(tert-butyl)[(5-chloro(2-thienyl))sulfonyl]amine was cooled to −78° C.and treated with 954 μL (1.5 mmol) of a 1.6 M solution of butyl lithiumin hexane. After 1 h, 159 mg (0.5 mmol) ofbis(phenylsulfonyl)fluoroamine was added and the solution was allowed towarm to 23° C. The reaction was quenched with 1 mL of sat. NH₄Cl,extracted 3 times with diethyl ether, dried (MgSO₄), concentrated invacuo to afford a quantitative yield (147 mg) of the desire product.¹⁹F-NMR (CDCl₃) δ (ppm): −113.4.

C. Synthesis ofN-[4-(2,4-dioxo(1,3-dihydroquinazolin-3-yl))phenyl]{[(5-chloro-3-fluoro(2-thienyl))sulfonyl]amino}carboxamide

A 19 mg-sample (0.07 mmol) of(tert-butyl)[(5-chloro-3-fluoro(2-thienyl))sulfonyl]amine was dissolvedin neat TFA and stirred for 1 h, concentrated in vacuo and used directlyin the next transformation. This sample was dissolved in 150 uL of DCMand 21 mg (0.084 mmol) of DSC was added followed by 21 μL (0.14 mmol) ofDBU. This solution was stirred for 18 h, 26 mg (0.07) of3-(4-aminophenyl)-1,3-dihydroquinazoline-2,4-dione trifluoroacetate saltwas added plus 150 μL of dry acetonitrile and refluxed for 2 h. Thismaterial was then purified on RP-HPLC to afford 11 mg (34%) of thedesired product. ES-MS: M+H+=495 (Cl).

Pharmaceutical Compositions and Methods of Treatment

A compound of formulae (I)–(VIII) according to the invention may beformulated into pharmaceutical compositions. Accordingly, the inventionalso relates to a pharmaceutical composition for preventing or treatingthrombosis in a mammal, particularly those pathological conditionsinvolving platelet aggregation, containing a therapeutically effectiveamount of a compound of formulae (I)–(VIII) or a pharmaceuticallyacceptable salt thereof, each as described above, and a pharmaceuticallyacceptable carrier or agent. Preferably, a pharmaceutical composition ofthe invention contains a compound of formulae (I)–(VIII), or a saltthereof, in an amount effective to inhibit platelet aggregation, morepreferably, ADP-dependent aggregation, in a mammal, in particular, ahuman. Pharmaceutically acceptable carriers or agents include thoseknown in the art and are described below.

Pharmaceutical compositions of the invention may be prepared by mixingthe compound of formulae (I)–(VIII) with a physiologically acceptablecarrier or agent. Pharmaceutical compositions of the invention mayfurther include excipients, stabilizers, diluents and the like and maybe provided in sustained release or timed release formulations.Acceptable carriers, agents, excipients, stablilizers, diluents and thelike for therapeutic use are well known in the pharmaceutical field, andare described, for example, in Remington's Pharmaceutical Sciences, MackPublishing Co., ed. A. R. Gennaro (1985). Such materials are nontoxic tothe recipients at the dosages and concentrations employed, and includebuffers such as phosphate, citrate, acetate and other organic acidsalts, antioxidants such as ascorbic acid, low molecular weight (lessthan about ten residues) peptides such as polyarginine, proteins, suchas serum albumin, gelatin, or immunoglobulins, hydrophilic polymers suchas polyvinylpyrrolidinone, amino acids such as glycine, glutamic acid,aspartic acid, or arginine, monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, mannoseor dextrins, chelating agents such as EDTA, sugar alcohols such asmannitol or sorbitol, counterions such as sodium and/or nonionicsurfactants such as TWEEN, or polyethyleneglycol.

Methods for preventing or treating thrombosis in a mammal embraced bythe invention administer a therapeutically effective amount of acompound of formulae (I)–(VIII) alone or as part of a pharmaceuticalcomposition of the invention as described above to a mammal, inparticular, a human. Compounds of formulae (I)–(VIII) and pharmaceuticalcompositions of the invention containing a compound of formulae(I)–(VIII) of the invention are suitable for use alone or as part of amulti-component treatment regimen for the prevention or treatment ofcardiovascular diseases, particularly those related to thrombosis. Forexample, a compound or pharmaceutical composition of the invention maybe used as a drug or therapeutic agent for any thrombosis, particularlya platelet-dependent thrombotic indication, including, but not limitedto, acute myocardial infarction, unstable angina, chronic stable angina,transient ischemic attacks, strokes, peripheral vascular disease,preeclampsia/eclampsia, deep venous thrombosis, embolism, disseminatedintravascular coagulation and thrombotic cytopenic purpura, thromboticand restenotic complications following invasive procedures, e.g.,angioplasty, carotid endarterectomy, post CABG (coronary artery bypassgraft) surgery, vascular graft surgery, stent placements and insertionof endovascular devices and protheses.

Compounds and pharmaceutical compositions of the invention may also beused as part of a multi-component treatment regimen in combination withother therapeutic or diagnostic agents in the prevention or treatment ofthrombosis in a mammal. In certain preferred embodiments, compounds orpharmaceutical compositions of the invention may be coadministered alongwith other compounds typically prescribed for these conditions accordingto generally accepted medical practice such as anticoagulant agents,thrombolytic agents, or other antithrombotics, including plateletaggregation inhibitors, tissue plasminogen activators, urokinase,prourokinase, streptokinase, heparin, aspirin, or warfarin.Coadministration may also allow for application of reduced doses of thethrombolytic agents and therefore minimize potential hemorrhagicside-effects. Compounds and pharmaceutical compositions of the inventionmay also act in a synergistic fashion to prevent reocclusion following asuccessful thrombolytic therapy and/or reduce the time to reperfusion.

The compounds and pharmaceutical compositions of the invention may beutilized in vivo, ordinarily in mammals such as primates, (e.g.,humans), sheep, horses, cattle, pigs, dogs, cats, rats and mice, or invitro. The biological properties, as defined above, of a compound or apharmaceutical composition of the invention can be readily characterizedby methods that are well known in the art such as, for example, by invivo studies to evaluate antithrombotic efficacy, and effects onhemostasis and hematological parameters.

Compounds and pharmaceutical compositions of the invention may be in theform of solutions or suspensions. In the management of thromboticdisorders the compounds or pharmaceutical compositions of the inventionmay also be in such forms as, for example, tablets, capsules or elixirsfor oral administration, suppositories, sterile solutions or suspensionsor injectable administration, and the like, or incorporated into shapedarticles. Subjects (typically mammalian) in need of treatment using thecompounds or pharmaceutical compositions of the invention may beadministered dosages that will provide optimal efficacy. The dose andmethod of administration will vary from subject to subject and bedependent upon such factors as the type of mammal being treated, itssex, weight, diet, concurrent medication, overall clinical condition,the particular compound of formulae (I)–(VIII) employed, the specificuse for which the compound or pharmaceutical composition is employed,and other factors which those skilled in the medical arts willrecognize.

Dosage formulations of compounds of formulae (I)–(VIII), orpharmaceutical compositions contain a compound of the invention, to beused for therapeutic administration must be sterile. Sterility isreadily accomplished by filtration through sterile membranes such as 0.2micron membranes, or by other conventional methods. Formulationstypically will be stored in a solid form, preferably in a lyophilizedform. While the preferred route of administration is orally, the dosageformulations of compounds of formulae (I)–(VIII) or pharmaceuticalcompositions of the invention may also be administered by injection,intravenously (bolus and/or infusion), subcutaneously, intramuscularly,colonically, rectally, nasally, transdermally or intraperitoneally. Avariety of dosage forms may be employed as well including, but notlimited to, suppositories, implanted pellets or small cylinders,aerosols, oral dosage formulations and topical formulations such asointments, drops and dermal patches. The compounds of formulae(I)–(VIII) and pharmaceutical compositions of the invention may also beincorporated into shapes and articles such as implants which may employinert materials such biodegradable polymers or synthetic silicones as,for example, SILASTIC, silicone rubber or other polymers commerciallyavailable. The compounds and pharmaceutical compositions of theinvention may also be administered in the form of liposome deliverysystems, such as small unilamellar vesicles, large unilamellar vesiclesand multilamellar vesicles. Liposomes can be formed from a variety oflipids, such as cholesterol, stearylamine or phosphatidylcholines.

Therapeutically effective dosages may be determined by either in vitroor in vivo methods. For each particular compound or pharmaceuticalcomposition of the invention, individual determinations may be made todetermine the optimal dosage required. The range of therapeuticallyeffective dosages will be influenced by the route of administration, thetherapeutic objectives and the condition of the patient. For injectionby hypodermic needle, it may be assumed the dosage is delivered into thebodily fluids. For other routes of administration, the absorptionefficiency must be individually determined for each compound by methodswell known in pharmacology. Accordingly, it may be necessary for thetherapist to titer the dosage and modify the route of administration asrequired to obtain the optimal therapeutic effect.

The determination of effective dosage levels, that is, the dosage levelsnecessary to achieve the desired result, i.e., platelet ADP receptorinhibition, will be readily determined by one skilled in the art.Typically, applications of a compound or pharmaceutical composition ofthe invention are commenced at lower dosage levels, with dosage levelsbeing increased until the desired effect is achieved. The compounds andcompositions of the invention may be administered orally in an effectiveamount within the dosage range of about 0.01 to 1000 mg/kg in a regimenof single or several divided daily doses. If a pharmaceuticallyacceptable carrier is used in a pharmaceutical composition of theinvention, typically, about 5 to 500 mg of a compound of formulae(I)–(VIII) is compounded with a pharmaceutically acceptable carrier ascalled for by accepted pharmaceutical practice including, but notlimited to, a physiologically acceptable vehicle, carrier, excipient,binder, preservative, stabilizer, dye, flavor, etc. The amount of activeingredient in these compositions is such that a suitable dosage in therange indicated is obtained.

Typical adjuvants which may be incorporated into tablets, capsules andthe like include, but are not limited to, binders such as acacia, cornstarch or gelatin, and excipients such as microcrystalline cellulose,disintegrating agents like corn starch or alginic acid, lubricants suchas magnesium stearate, sweetening agents such as sucrose or lactose, orflavoring agents. When a dosage form is a capsule, in addition to theabove materials it may also contain liquid carriers such as water,saline, or a fatty oil. Other materials of various types may be used ascoatings or as modifiers of the physical form of the dosage unit.Sterile compositions for injection can be formulated according toconventional pharmaceutical practice. For example, dissolution orsuspension of the active compound in a vehicle such as an oil or asynthetic fatty vehicle like ethyl oleate, or into a liposome may bedesired. Buffers, preservatives, antioxidants and the like can beincorporated according to accepted pharmaceutical practice.

Pharmacological Assays

The pharmacological activity of each of the compounds according to theinvention is determined by the following in vitro assays:

I. Inhibition of ADP-Mediated Platelet Aggregation in vitro

The effect of testing the compound according to the invention onADP-induced human platelet aggregation is preferably assessed in 96-wellmicrotiter assay (see generally the procedures in Jantzen, H. M. et al.(1999) Thromb. Hemost. 81:111–117). Human venous blood is collected fromhealthy, drug-free volunteers into ACD (85 mM sodium citrate, 111 mMglucose, 71.4 mM citric acid) containing PGI₂ (1.25 ml ACD containing1.6 μM PGI₂/10 ml blood; PGI₂ was from Sigma, St. Louis, Mo.).Platelet-rich plasma (PRP) is prepared by centrifugation at 160×g for 20minutes at room temperature. Washed platelets are prepared bycentrifuging PRP for 10 minutes at 730 g and resuspending the plateletpellet in CGS (13 mM sodium citrate, 30 mM glucose, 120 mM NaCl; 2 mlCGS/10 mil original blood volume) containing 1 U/ml apyrase (grade V,Sigma, St. Louis, Mo.). After incubation at 37° C. for 15 minutes, theplatelets are collected by centrifugation at 730 g for 10 minutes andresuspended at a concentration of 3×10⁸ platelets/ml in Hepes-Tyrode'sbuffer (10 mM Hepes, 138 mM NaCl, 5.5 mM glucose, 2.9 mM KCl, 12 mMNaHCO₃, pH 7.4) containing 0.1% bovine serum albumin, 1 mM CaCl₂ and 1mM MgCl₂. This platelet suspension is kept >45 minutes at 37° C. beforeuse in aggregation assays.

Inhibition of ADP-dependent aggregation is preferably determined in96-well flat-bottom microtiter plates using a microtiter plate shakerand plate reader similar to the procedure described by Frantantoni etal., Am. J. Clin. Pathol. 94, 613 (1990). All steps are performed atroom temperature. The total reaction volume of 0.2 mil/well includes inHepes-Tyrodes buffer/0.1% BSA: 4.5×10⁷ apyrase-washed platelets, 0.5mg/ml human fibrinogen (American Diagnostica, Inc., Greenwich, Conn.),serial dilutions of test compounds (buffer for control wells) in 0.6%DMSO. After about 5 minutes preincubation at room temperature, ADP isadded to a final concentration of 2:M which induces submaximalaggregation. Buffer is added instead of ADP to one set of control wells(ADP⁻ control). The OD of the samples is then determined at 490 nm usinga microtiter plate reader (Softmax, Molecular Devices, Menlo Park,Calif.) resulting in the 0 minute reading. The plates are then agitatedfor 5 min on a microtiter plate shaker and the 5 minute reading isobtained in the plate reader. Aggregation is calculated from thedecrease of OD at 490 nm at t=5 minutes compared to t=0 minutes and isexpressed as % of the decrease in the ADP control samples aftercorrecting for changes in the unaggregated control samples.

II. Inhibition of [³H]12-MeS-ADP Binding to Platelets

Having first determined that the compounds according to the inventioninhibit ADP-dependent platelet aggregation with the above assay, asecond assay is used to determine whether such inhibition is mediated byinteraction with platelet ADP receptors. Utilizing the second assay thepotency of inhibition of such compounds with respect to [³H]2-MeS-ADPbinding to whole platelets is determined. [³H]2-MeS-ADP bindingexperiments are routinely performed with outdated human plateletscollected by standard procedures at hospital blood banks. Apyrase-washedoutdated platelets are prepared as follows (all steps at roomtemperature, if not indicated otherwise):

Outdated platelet suspensions are diluted with 1 volume of CGS andplatelets pelleted by centrifugation at 1900×g for 45 minutes. Plateletpellets are resuspended at 3–6×10⁹ platelets /ml in CGS containing 1U/ml apyrase (grade V, Sigma, St. Louis, Mo.) and incubated for 15minutes at 37° C. After centrifugation at 730×g for 20 minutes, pelletsare resuspended in Hepes-Tyrode's buffer containing 0.1% BSA (Sigma, St.Louis, Mo.) at a concentration of 6.66×10⁸ platelets/ml. Bindingexperiments are performed after >45 minutes resting of the platelets.

Alternatively, binding experiments are performed with fresh humanplatelets prepared as described in I.(Inhibition of ADP-MediatedPlatelet Aggregation in vitro), except that platelets are resuspended inHepes-Tyrode's buffer containing 0.1% BSA (Sigma, St. Louis, Mo.) at aconcentration of 6.66×10⁸ platelets/ml. Very similar results areobtained with fresh and outdated platelets.

A platelet ADP receptor binding assay using the tritiated potent agonistligand [³H]2-MeS-ADP (Jantzen, H. M. et al. (1999) Thromb. Hemost.81:111–117) has been adapted to the 96-well microtiter format. In anassay volume of 0.2 ml Hepes-Tyrode's buffer with 0.1% BSA and 0.6%DMSO, 1×10⁸ apyrase-washed platelets are preincubated in 96-well flatbottom microtiter plates for 5 minutes with serial dilutions of testcompounds before addition of 1 nM [³H]2-MeS-ADP([³H]2-methylthioadenosine-5′-diphosphate, ammonium salt; specificactivity 48–49 Ci/mmole, obtained by custom synthesis from Amersham LifeScience, Inc., Arlington Heights, Ill., or NEN Life Science Products,Boston, Mass.). Total binding is determined in the absence of testcompounds. Samples for nonspecific binding may contain 10⁻⁵ M unlabelled2-MeS-ADP (RBI, Natick, Mass.). After incubation for 15 minutes at roomtemperature, unbound radioligand is separated by rapid filtration andtwo washes with cold (4–8° C.) Binding Wash Buffer (10 mM Hepes pH 7.4,138 mM NaCl) using a 96-well cell harvester (Minidisc 96, SkatronInstruments, Sterling, Va.) and 8×12 GF/C glassfiber filtermats (PrintedFiltermat A, for 1450 Microbeta, Wallac Inc., Gaithersburg, Md.). Theplatelet-bound radioactivity on the filtermats is determined in ascintillation counter (Microbeta 1450, Wallac Inc., Gaithersburg, Md.).Specific binding is determined by subtraction of nonspecific bindingfrom total binding, and specific binding in the presence of testcompounds is expressed as % of specific binding in the absence of testcompounds dilutions.

It should be understood that the foregoing discussion, embodiments andexamples merely present a detailed description of certain preferredembodiments. It will be apparent to those of ordinary skill in the artthat various modifications and equivalents can be made without departingfrom the spirit and scope of the invention. All the patents, journalarticles and other documents discussed or cited above are hereinincorporated by reference.

1. A compound selected from the group consisting of formula (I) andformula (II):

wherein: A is thienyl optionally substituted with one, two or threesubstituents independently selected from the group consisting of loweralkoxy, lower alkyl, loweralkylamino, hydroxy, halogen, cyano, hydroxyl,mercapto, nitro, thioalkoxy, carboxaldehyde, carboxyl, carboalkoxy andcarboxamide; W is 1,4-phenylene optionally substituted with one, two orthree substituents independently selected from the group consisting oflower alkoxy, lower alkyl, lower alkylamino, hydroxy, halogen, cyano,hydroxyl, mercapto, nitro, thioalkoxy, carboxaldehyde, carboxyl,carboalkoxy and carboxamide; E is selected from the group consisting ofH, —C₁–C₈ alkyl, polyhaloalkyl, —C₃₋₈-cycloalkyl, aryl optionallysubstituted with one, two or three substituents independently selectedfrom the group consisting of lower alkoxy, lower alkyl, loweralkylamino, hydroxy, halogen, cyano, hydroxyl, mercapto, nitro,thioalkoxy, carboxaldehyde, carboxyl, carboalkoxy and carboxamide D is

wherein n is an integer from 0–4, X is in each case a memberindependently selected from the group consisting of: halogen,polyhaloalkyl, —OR³, —SR³, —CN, —NO₂, —SO₂R³, —C₁₋₁₀-alkyl,C₃₋₈-cycloalkyl, aryl, aryl-substituted by 1–4 R³ groups, amino,amino-C₁₋₈-alkyl, C₁₋₃-acylamino, C₁₋₃-acylamino-C₁₋₈-alkyl,C₁₋₆-alkylamino, C₁₋₆-alkylamino C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆dialkylamino C₁₋₈ alkyl, C₁₋₆ alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl,carboxy-C₁₋₆-alkyl, C₁₋₃alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl,carboxy C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl, and a 5 to 10membered fused or non-fused aromatic or nonaromatic heterocyclic ringsystem, having 1 to 4 heteroatoms independently selected from N, O, andS, with the proviso that the carbon and nitrogen atoms, when present inthe heterocyclic ring system, are unsubstituted, mono- or di-substitutedindependently with 0–2 R⁴ groups, wherein R³ and R⁴ are eachindependently selected from the group consisting of: hydrogen, halogen,—CN, —NO₂, —C₁₋₁₀ alkyl, C₃₋₈-cycloalkyl, aryl, amino, amino-C₁₋₈-alkyl,C₁₋₃-acylamino, C₁₋₃-acylamino-C₁₋₈-alkyl, C₁₋₆-alkylamino,C₁₋₆-alkylamino C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈alkyl, C₁₋₆ alkoxy, C₁₋₆ alkoxy-C₁₋₆-alkyl, carboxy-C₁₋₆-alkyl,C₁₋₃-alkoxycarbonyl, C₁₋₃-alkoxycarbonyl-C₁₋₆-alkyl,carboxy-C₁₋₆-alkyloxy, hydroxy, hydroxy-C₁₋₆-alkyl, -thio andthio-C₁₋₆-alkyl; Y is selected from the group consisting of O, S, N—OR⁵,and NR⁵, wherein R⁵ is selected from the group consisting of: H, C₁₋₁₀alkyl, C₃₋₈-cycloalkyl, NR², and CN; and or pharmaceutically acceptablesalts thereof.
 2. A compound according to claim 1, wherein: A isselected from the group consisting of:

Y is selected from the group consisting of O, S, N—OR⁵ and NR⁵. E isselected from the group consisting of H and C₁₋₈ alkyl. W is selectedfrom the group consisting of:


3. A compound according to claim 1, wherein: D is selected from thegroup consisting of:


4. A compound of claim 1, selected from the group consisting of:


5. A pharmaceutical composition comprising a therapeutically effectiveamount of a compound according to claim 1, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier.
 6. Amethod for preventing or treating thrombosis in a mammal comprising thestep of administering to a mammal a therapeutically effective amount ofa compound of claim 1 or a pharmaceutically acceptable salt thereof. 7.A method of claim 6, wherein said mammal is a human.
 8. A method ofclaim 6, wherein said mammal is prone to or suffers from acardiovascular disease selected from the group consisting of acutemyocardial infarction, unstable angina, chronic stable angina, transientischemic attacks, strokes, peripheral vascular disease,preeclampsia/eclampsia, deep venous thrombosis, embolism, disseminatedintravascular coagulation and thrombotic cytopenic purpura, thormboticand restenotic complications following invasive procedures resultingfrom angioplasty, carotid endarterectomy, post CABG (Coronary arterylbypass graft) surgery, vascular graft surgery, stent placements andinsertion of endovascular devices and prothesis.
 9. A compound of claim1, having a formula selected from the group consisting of:

wherein: n is an integer from 0–4; X is selected from the groupconsisting of Cl, OMe and NHMe or taken together with the ring to whichthey are attached to form atetrahydro-triaza-cyclopentanaphthalene-dione moiety; W is selected fromthe group consisting of:

Y is O; and A is:


10. A compound of claim 1, having the following formula:

wherein: Y is O; and A is


11. A compound of claim 1 wherein A is thienyl.
 12. A compound of claim1 wherein Y is O.
 13. A compound of claim 1 wherein E is hydrogen.
 14. Acompound of claim 1, having the following formula:


15. A compound of claim 1, having the following formula:


16. A compound of claim 1, having the following formula:


17. A compound of claim 1, having the following formula:


18. A compound of claim 1, having the following formula:


19. A compound of claim 1, having the following formula:


20. A compound of claim 1, having the following formula: